_:b547722553 . _:b547722552 . _:b547722127 . _:b547722555 . _:b29675228 . _:b547722554 . _:b547722565 . . . _:b547722564 . . _:b547722567 . . _:b547722448 . _:b547722347 . _:b547722566 . _:b547722561 . _:b547722560 . _:b547722563 . _:b547722562 . . _:b547722573 . . _:b547722240 . _:b547722572 . _:b547722636 . . _:b547722575 . _:b29675252 . _:b547722170 . _:b547722480 . _:b547722574 . _:b547722569 . _:b547722568 . _:b547722675 . . _:b547722571 . . . _:b547722371 . _:b547722570 . _:b547722581 . . . . _:b547722580 . _:b547722583 . _:b29675295 "However, it should be noted that fetal liver cell repopulation is also low in those mouse models [>>53<<]. Genetic modification of human liver cells for enhanced survival, repopulation and differentiation coupled with mouse Fas agonist (Jo2) [34] and/or the AFC8 murine liver damage system [18] could further enhance human liver repopulation." . _:b547722582 . . _:b547722577 . _:b547722385 . _:b547722576 . . . . _:b547722579 . _:b29675239 . _:b547722428 . _:b547722578 . _:b547722294 . _:b547722589 . _:b547722588 . _:b547722377 . _:b547722591 . _:b547722590 . _:b29675299 "In chronically infected patients, immune and inflammatory responses against HBV are implicated as the major mediators of liver diseases [>>54<<], [55]. Chronic HBV infection in the liver of A2/NSG/Fas-hu mice was associated with significant human leukocyte infiltration, leading to human hepatic stellate cell activation and human liver fibrosis." . . _:b547722585 . . _:b547722584 . . _:b547722587 . . _:b547722586 . _:b547722206 . _:b547722597 . _:b547722596 . . _:b547722599 . _:b547722590 . _:b547722598 . _:b547722218 . _:b547722593 . _:b547722592 . _:b547722595 . _:b547722594 . _:b547722411 . _:b547722228 . _:b547722605 . _:b547722220 . _:b547722604 . _:b547722660 . . _:b547722607 . _:b547722611 . _:b547722606 . _:b547722601 . _:b29675224 "To overcome the limitations associated with current chimeric human-murine liver mouse models, we have recently developed a humanized mouse model with both human immune system and liver cells (AFC8-hu HSC/Hep mice) [>>18<<], [19]. AFC8-hu HSC/Hep mice can support HCV infection in the liver and generate human T-cell response to HCV. Additionally, HCV infection induces liver inflammation and fibrosis, correlated with activation of human hepatic stellate cells" . _:b547722600 . _:b547722603 . _:b547722602 . . . _:b547722111 . _:b547722613 . _:b547722249 . _:b547722612 . _:b547722615 . _:b547722293 . _:b547722340 . _:b29675255 . _:b547722614 . . _:b547722609 . _:b547722608 . _:b547722359 . _:b547722611 . _:b547722610 . _:b547722621 . _:b547722531 . _:b547722676 . _:b547722677 . _:b547722412 . _:b547722620 . . _:b547722672 . _:b547722200 . _:b547722673 . _:b547722623 . _:b547722674 . _:b547722675 . _:b547722668 . _:b547722669 . _:b547722622 . _:b547722670 . _:b547722671 . _:b547722664 . _:b547722665 . _:b547722617 . . _:b547722666 . _:b547722667 . _:b547722660 . _:b547722661 . _:b547722616 . _:b547722662 . _:b547722255 . _:b547722663 . _:b547722364 . _:b547722656 . _:b547722657 . _:b547722619 . _:b547722658 . _:b547722659 . . _:b547722652 . _:b547722653 . _:b547722208 . _:b547722618 . _:b547722654 . _:b547722185 . _:b547722655 . _:b547722378 . _:b547722648 . _:b547722649 . _:b547722629 . _:b547722650 . _:b547722651 . _:b547722644 . _:b547722645 . _:b547722628 . _:b547722646 . _:b547722647 . _:b547722640 . . _:b547722631 . _:b547722641 . . _:b547722642 . _:b547722643 . _:b547722636 . _:b547722637 . _:b547722630 . _:b547722638 . _:b547722639 . _:b547722512 . _:b547722632 . . _:b547722633 . _:b29675254 . _:b547722625 . _:b547722634 . _:b547722635 . _:b547722628 . _:b547722629 . . _:b547722624 . _:b547722630 . _:b547722631 . _:b547722624 . _:b547722625 . _:b547722627 . _:b547722626 . _:b547722627 . . _:b547722626 . _:b547722637 . _:b547722636 . . _:b29675271 "disease accounts for the vast majority of HBV associated morbidity/mortality, acute HBV infection occasionally results in accelerated liver disease and liver failure with subsequent mortality in the absence of liver transplantation [>>8<<]. Analysis of liver gene expression profile in acute HBV-induced liver failure patients also showed increased macrophage infiltration (CD68 upregulation), up-regulation of M2-like macrophage genes (IL10RA - Interleukin 10 receptor alpha" . . _:b547722639 . . _:b547722638 . _:b547722633 . _:b29675211 "liver disease is predominately a chronic disease, requiring decades of chronic infection and liver inflammation [5], [6], [7], HBV infection occasionally results in accelerated liver disease and liver failure during acute infection [>>8<<], [9]. The development of preventive vaccines and therapeutics using chimpanzees and surrogate hepatitis virus-small animal models has played a significant role in preventing new infections and controlling HBV-induced liver diseases." . _:b29675221 "Additionally, the fumarylacetoacetate hydrolase (Fah)-Rag2-\u03B3C-null mice also allow human hepatocytes engraftment and HCV infection [15], [16], [>>17<<]. However, these human-murine chimeric liver models lack a functional human immune system, thus it is not possible to study host immune response and hepatitis virus-induced immunopathology [14], [17]. To overcome the limitations associated" . _:b29675268 . _:b547722632 . . _:b29675269 . _:b547722635 . _:b547722243 . _:b29675270 . _:b547722634 . _:b29675277 . . _:b29675271 . _:b547722645 . _:b29675273 "- alternative macrophage activation-associated CC chemokine-1, IL10, B7-H4) and down-regulation or no change of M1-like macrophage genes (TNF\u03B1, iNOS, IL12p40) in HBV infected patients compared to healthy controls (Figure S8, S9) [8], [>>9<<]. Furthermore, liver gene expression profile analysis showed M2-like macrophage gene expression profile (CD68high, CD163high, AMAC1high, iNOSlow, TNF\u03B1low) is associated with upregulation of tissue fibrosis (COL1A1high, TIMP1high) and" . _:b29675264 . _:b547722644 . _:b29675265 . _:b547722647 . _:b29675266 . _:b547722302 . . _:b547722646 . _:b29675267 . _:b547722641 . _:b547722616 . _:b29675276 . _:b547722640 . _:b547722424 . _:b547722631 . _:b29675277 . _:b547722421 . . _:b547722643 . _:b29675278 . _:b547722556 . _:b547722557 . _:b547722642 . . _:b29675279 . _:b547722558 . . _:b547722559 . _:b547722552 . . _:b547722653 . _:b547722553 . _:b547722554 . _:b29675272 . _:b547722555 . _:b547722548 . _:b547722549 . _:b547722652 . _:b547722550 . _:b29675273 . _:b547722549 . _:b547722551 . _:b547722544 . _:b547722545 . _:b547722655 . _:b547722546 . _:b29675274 . _:b547722547 . _:b547722162 . _:b547722540 . _:b547722541 . _:b547722654 . _:b547722542 . _:b29675275 . _:b547722543 . . _:b547722536 . _:b547722537 . _:b547722649 . _:b547722276 . _:b29675284 . _:b547722538 . . _:b547722539 . _:b547722532 . _:b29675277 "showed M2-like macrophage gene expression profile (CD68high, CD163high, AMAC1high, iNOSlow, TNF\u03B1low) is associated with upregulation of tissue fibrosis (COL1A1high, TIMP1high) and damage markers (HMGB1high [42], [43], [44], S100A9high [>>45<<], [46]) (Figure S9) [8], [9]." . _:b547722533 . _:b547722648 . _:b547722534 . _:b29675285 . _:b547722535 . _:b547722146 . _:b547722528 . . _:b547722529 . _:b547722651 . _:b547722530 . _:b547722224 . _:b29675286 . _:b547722531 . _:b547722524 . _:b547722525 . _:b547722650 . _:b547722526 . _:b29675287 . _:b547722527 . _:b547722520 . _:b547722521 . _:b29675226 . _:b547722661 . _:b547722522 . _:b29675280 . _:b547722523 . _:b547722516 . _:b547722517 . _:b547722660 . _:b547722518 . _:b29675281 . _:b29675254 "RNA was isolated from tissue following manufacturer's recommended procedures and qPCR was performed using species-specific published [>>18<<], [61] or NCBI primer blast generated primers and the SYBR Green method, following manufacturer's recommended procedures (Thermo Scientific)." . _:b29675236 "a humanized mouse model by injecting human liver progenitor cells (Hep) and CD34+ human hematopoietic stem cells (HSC) directly into the liver of newborn A2/NSG (HLA-A2 transgenic NOD scid IL2 receptor gamma chain knockout mice [28], [>>29<<], [30]). The A2/NSG mouse lacks NK cells and T/B-lymphocytes. They support efficient development of a functional human immune system after injecting CD34+ human hematopoietic stem cells (HSC) into the liver of newborn mice [30], [31]." . _:b547722316 . _:b547722519 . _:b547722512 . _:b547722513 . _:b29675224 . _:b547722663 . _:b547722514 . _:b29675282 . _:b547722513 . _:b547722630 . . _:b547722515 . _:b547722508 . _:b547722509 . _:b547722662 . _:b547722510 . _:b29675283 . _:b547722511 . . _:b547722504 . _:b547722505 . _:b547722657 . _:b547722506 . _:b29675292 . _:b547722507 . _:b547722500 . _:b29675214 . _:b547722501 . _:b547722656 . _:b547722502 . _:b29675293 . _:b547722503 . _:b547722496 . _:b547722497 . _:b547722659 . _:b547722498 . _:b29675294 . _:b547722499 . _:b547722620 . _:b547722621 . _:b547722658 . _:b547722622 . _:b29675295 . _:b547722623 . _:b547722616 . _:b547722617 . _:b547722669 . _:b547722618 . _:b29675288 . _:b547722619 . _:b547722229 . _:b547722612 . _:b547722613 . _:b547722668 . _:b547722614 . _:b29675289 . _:b547722615 . _:b547722608 . _:b547722420 . _:b547722671 . _:b547722609 . _:b547722610 . _:b29675290 . _:b547722611 . _:b547722372 . _:b547722604 . _:b547722605 . _:b547722670 . _:b547722606 . _:b29675291 . _:b547722607 . _:b547722600 . _:b547722601 . _:b547722665 . . _:b29675300 . _:b547722602 . _:b547722552 . _:b547722603 . _:b547722596 . _:b547722597 . _:b547722664 . . _:b29675301 . _:b547722598 . _:b547722599 . _:b547722592 . _:b547722593 . _:b547722667 . _:b547722594 . _:b29675302 . _:b547722595 . _:b547722588 . _:b547722589 . _:b547722666 . _:b547722590 . _:b29675303 . _:b547722149 . _:b547722368 . _:b547722591 . _:b547722584 . _:b547722585 . _:b547722677 . _:b547722586 . _:b29675296 . _:b547722587 . _:b547722580 . . _:b547722676 . _:b547722581 . _:b547722540 . _:b547722582 . _:b29675297 . _:b547722414 . _:b547722583 . _:b547722576 . _:b547722577 . _:b547722578 . _:b29675298 . _:b547722579 . _:b547722572 . _:b547722573 . _:b547722574 . _:b29675299 . _:b547722575 . _:b29675242 "To promote human liver cell repopulation, A2/NSG-hu HSC/Hep mice were treated with a murine specific anti-Fas agonistic antibody (Jo2) [32], [>>33<<], [34], [35], [36]." . _:b547722568 . _:b547722569 . _:b547722673 . _:b547722570 . _:b29675308 . _:b547722541 . _:b547722571 . _:b547722231 . . _:b547722564 . _:b547722565 . _:b547722672 . _:b547722566 . _:b29675309 . _:b547722567 . . _:b547722560 . _:b547722561 . _:b547722675 . _:b547722562 . _:b547722563 . _:b547722428 . _:b547722429 . _:b547722674 . _:b29675204 "introduction" . _:b547722196 . _:b547722430 . . _:b547722431 . _:b29675204 _:b29675236 . _:b547722424 . _:b29675204 _:b29675237 . _:b547722425 . _:b29675204 _:b29675238 . _:b29675304 . _:b547722426 . _:b29675204 _:b29675239 . _:b547722427 . _:b29675204 _:b29675232 . _:b547722420 . _:b29675204 _:b29675233 . _:b547722421 . _:b29675204 _:b29675234 . _:b29675305 . _:b547722422 . _:b29675204 _:b29675235 . _:b547722423 . _:b29675204 _:b29675244 . _:b547722416 . _:b29675204 _:b29675245 . _:b547722417 . _:b547722418 . _:b29675306 . _:b547722419 . . _:b547722116 . _:b547722412 . _:b29675204 _:b29675240 . _:b29675204 _:b29675241 . . _:b547722413 . _:b29675204 _:b29675242 . _:b29675307 . _:b547722414 . . _:b29675204 _:b29675243 . _:b547722415 . _:b29675204 _:b29675220 . _:b547722408 . _:b29675204 _:b29675221 . _:b547722409 . _:b29675204 _:b29675222 . _:b547722410 . _:b29675204 _:b29675223 . _:b547722411 . _:b29675204 _:b29675216 . _:b547722404 . _:b29675204 _:b29675217 . _:b547722405 . _:b29675204 _:b29675218 . _:b547722406 . _:b29675204 _:b29675219 . _:b547722407 . . _:b547722400 . _:b29675204 _:b29675228 . _:b29675204 _:b29675229 . _:b547722401 . _:b29675204 _:b29675230 . _:b547722156 . _:b547722402 . _:b29675204 _:b29675231 . _:b547722403 . _:b29675204 _:b29675224 . _:b547722396 . _:b29675204 _:b29675225 . _:b547722397 . _:b29675204 _:b29675226 . _:b547722398 . _:b29675204 _:b29675227 . _:b547722399 . _:b547722392 . _:b29675266 "However, antigen-specific IgG response was detected in only two of eight mice and at very low levels as reported in other human B cell studies with humanized mice [>>37<<]. To characterize anti-HBV human T cell immune response, HLA-A2 donor derived-leukocytes from the spleen and lymph nodes of mock- or HBV-infected humanized animals were collected and stimulated in vitro with PHA or A2-restricted HBV" . _:b547722401 . _:b547722393 . _:b29675204 _:b29675205 . _:b29675204 _:b29675206 . _:b547722394 . _:b29675204 _:b29675207 . _:b547722395 . _:b547722388 . _:b547722464 . _:b547722389 . _:b547722390 . _:b547722391 . _:b29675204 _:b29675212 . _:b547722384 . _:b29675204 _:b29675213 . _:b547722385 . _:b29675204 _:b29675214 . _:b547722386 . . _:b29675204 _:b29675215 . _:b547722387 . _:b29675204 _:b29675208 . _:b547722380 . _:b29675204 _:b29675209 . _:b547722381 . _:b29675204 _:b29675210 . _:b547722382 . _:b547722635 . _:b29675204 _:b29675211 . _:b547722383 . _:b547722376 . _:b29675306 "micronutrients to sites of wound repair; however, during chronic infection, M2-like macrophages promote tissue fibrosis, neoplasia and impair Th1 response thus promoting pathogen persistence and associated tissue pathology [>>40<<]. We report that liver inflammation and immune impairment in chronic HBV infected humanized mice livers was associated with M2-like macrophages, which also localized to fibrotic regions." . _:b547722377 . _:b547722378 . _:b547722175 . _:b547722379 . _:b547722372 . _:b547722481 . _:b547722373 . _:b547722374 . . _:b547722375 . _:b547722368 . _:b547722369 . _:b547722370 . _:b547722371 . _:b547722492 . . _:b547722493 . _:b547722494 . _:b547722495 . _:b547722488 . . _:b547722489 . _:b547722490 . _:b547722518 . _:b547722491 . _:b547722484 . _:b547722485 . _:b547722486 . _:b547722487 . _:b547722480 . _:b547722481 . _:b547722625 . _:b547722242 . _:b547722602 . _:b547722482 . _:b547722483 . _:b547722476 . . _:b547722477 . . _:b547722478 . _:b547722479 . _:b547722669 . _:b547722472 . _:b547722473 . _:b547722474 . _:b547722475 . _:b547722468 . _:b547722469 . _:b547722470 . _:b547722471 . _:b547722464 . . _:b547722435 . _:b547722465 . _:b547722466 . _:b547722467 . _:b547722460 . _:b547722461 . . _:b547722462 . _:b547722463 . _:b547722456 . _:b547722457 . _:b547722458 . _:b547722459 . _:b547722452 . _:b547722453 . _:b547722454 . _:b547722455 . _:b547722448 . _:b547722674 . _:b547722449 . _:b547722450 . _:b547722451 . _:b547722444 . _:b547722445 . _:b547722446 . _:b547722447 . . _:b547722440 . _:b547722441 . _:b547722442 . _:b547722443 . _:b547722436 . . _:b547722437 . _:b547722438 . . _:b547722439 . _:b547722623 . _:b547722432 . _:b547722433 . _:b29675282 "Results from several cell culture studies have suggested that HBV can modulate monocyte activation resulting in induction of M2 associated cytokines and inhibition of M1 associated cytokines [20], [>>22<<], [47], [48], [49]." . _:b547722434 . _:b547722435 . _:b547722300 . _:b547722301 . _:b547722302 . _:b547722303 . _:b547722296 . _:b547722297 . _:b547722298 . _:b547722299 . _:b547722292 . _:b547722293 . . _:b547722294 . _:b547722295 . _:b547722288 . _:b547722445 . _:b547722289 . _:b547722290 . _:b547722291 . _:b547722284 . _:b547722285 . _:b547722286 . _:b547722287 . _:b29675246 _:b29675260 . _:b547722280 . . _:b29675246 _:b29675261 . _:b547722281 . _:b547722432 . _:b29675246 _:b29675262 . _:b547722282 . _:b29675246 _:b29675263 . _:b547722283 . . _:b547722276 . _:b29675246 _:b29675256 . _:b29675246 _:b29675257 . _:b547722277 . _:b29675246 _:b29675258 . _:b547722278 . _:b29675246 _:b29675259 . _:b547722279 . _:b547722172 . . _:b29675246 _:b29675252 . _:b547722272 . _:b29675246 _:b29675253 . _:b547722273 . _:b29675246 _:b29675254 . _:b547722274 . _:b29675246 _:b29675255 . _:b547722275 . _:b29675246 _:b29675248 . _:b547722268 . _:b29675246 _:b29675249 . _:b547722269 . _:b29675246 _:b29675250 . _:b29675288 . _:b547722270 . . _:b547722271 . _:b547722647 . _:b29675246 _:b29675251 . _:b547722264 . _:b547722265 . _:b547722266 . _:b29675246 _:b29675247 . _:b547722267 . _:b547722260 . _:b29675270 . _:b547722261 . _:b547722262 . _:b547722263 . _:b547722256 . _:b547722257 . _:b547722258 . . _:b547722259 . _:b547722252 . _:b547722253 . _:b547722237 . _:b547722254 . _:b547722255 . _:b547722248 . _:b547722249 . _:b547722250 . . _:b547722587 . _:b547722251 . _:b547722244 . _:b547722245 . _:b547722246 . _:b547722247 . _:b547722240 . . _:b547722241 . _:b547722492 . _:b547722242 . _:b547722243 . _:b547722364 . _:b547722365 . . _:b547722366 . _:b547722261 . _:b547722367 . _:b547722396 . _:b547722360 . _:b547722361 . _:b547722362 . _:b547722215 . _:b547722363 . _:b547722356 . . _:b547722357 . _:b547722643 . _:b547722358 . _:b547722359 . _:b547722352 . _:b547722353 . _:b547722354 . _:b547722355 . _:b547722348 . _:b547722349 . _:b547722350 . _:b547722351 . _:b547722344 . _:b29675208 . . _:b547722345 . _:b547722346 . _:b29675261 "All patients were diagnosed according to our previously described criteria [64], [>>65<<] and had not received any antiviral therapies or immunosuppressive drugs within six months before sampling." . _:b547722347 . _:b547722340 . _:b547722341 . _:b547722342 . _:b547722343 . _:b547722336 . _:b547722337 . _:b547722338 . . _:b547722339 . _:b547722332 . _:b547722333 . _:b547722618 . _:b547722334 . _:b547722335 . _:b547722328 . _:b547722329 . _:b547722330 . . _:b547722331 . _:b547722324 . _:b547722325 . _:b547722326 . _:b547722483 . _:b29675212 "disease is predominately a chronic disease, requiring decades of chronic infection and liver inflammation [5], [6], [7], HBV infection occasionally results in accelerated liver disease and liver failure during acute infection [8], [>>9<<]. The development of preventive vaccines and therapeutics using chimpanzees and surrogate hepatitis virus-small animal models has played a significant role in preventing new infections and controlling HBV-induced liver diseases." . _:b547722327 . _:b547722320 . _:b547722321 . _:b547722322 . _:b547722323 . _:b547722316 . _:b547722317 . _:b547722318 . _:b547722319 . _:b547722312 . _:b547722313 . . _:b547722314 . _:b547722315 . _:b547722308 . _:b547722309 . _:b547722310 . _:b547722311 . _:b547722304 . _:b547722305 . _:b547722306 . _:b547722353 . _:b547722307 . . _:b547722172 . _:b547722173 . _:b547722174 . _:b547722175 . _:b547722168 . _:b547722169 . _:b547722170 . . _:b547722171 . _:b547722408 . _:b547722164 . _:b547722165 . _:b547722166 . _:b547722593 . _:b547722167 . . _:b547722160 . _:b547722161 . _:b547722162 . _:b547722163 . _:b547722156 . . . _:b547722157 . _:b547722158 . . _:b547722159 . _:b547722152 . _:b547722153 . _:b547722154 . _:b547722155 . . _:b547722142 . _:b547722148 . _:b547722149 . _:b547722356 . . _:b547722150 . _:b547722151 . _:b547722144 . _:b29675222 . _:b547722145 . _:b547722146 . _:b547722278 . _:b547722147 . _:b547722140 . _:b547722351 . _:b547722141 . _:b547722142 . _:b547722143 . _:b547722136 . _:b547722137 . _:b547722245 . _:b547722138 . . _:b547722139 . _:b547722132 . _:b29675217 . _:b547722133 . _:b547722244 . _:b547722134 . _:b547722135 . _:b547722128 . _:b29675218 . . _:b547722247 . _:b547722129 . _:b547722130 . _:b547722131 . _:b547722124 . _:b547722125 . _:b547722246 . _:b547722126 . _:b547722475 . _:b547722127 . _:b547722120 . _:b547722121 . _:b29675272 . _:b547722241 . _:b547722122 . _:b547722123 . _:b547722116 . _:b547722117 . _:b547722240 . . _:b547722118 . _:b547722119 . _:b547722112 . . _:b547722243 . _:b547722113 . _:b547722114 . . _:b547722115 . _:b547722236 . _:b547722237 . _:b547722242 . _:b547722238 . . _:b547722239 . _:b547722627 . _:b547722232 . _:b547722321 . _:b547722274 . _:b547722253 . _:b547722233 . _:b547722234 . _:b29675301 "and associated tissue fibrosis and damage; with M1 polarized macrophages promoting anti-virus Th1 immune response and pathogen clearance, while M2 polarized macrophages impair Th1 immune response and promoting tissue remodeling [>>23<<], [24], [25], [26], [27]." . _:b547722235 . _:b547722247 . _:b547722228 . _:b547722229 . _:b29675279 . _:b547722252 . _:b547722230 . _:b547722231 . _:b547722224 . _:b547722225 . _:b547722255 . _:b547722226 . _:b547722118 . _:b29675290 "model enables inducible depletion of murine hepatocytes through the Fas apoptotic signaling pathway, resulting in elevated human liver repopulation in mice transplanted with human liver progenitor and hematopoietic stem cells [32], [>>33<<], [34], [36], [51]. Additionally, the A2/NSG background permits highly efficient engraftment and development of human xenografts including human hematopoietic stem cells compared to current immunodeficient mouse models [52]." . _:b547722227 . _:b547722220 . _:b547722221 . _:b547722254 . _:b547722222 . _:b547722223 . _:b547722216 . _:b547722201 . _:b547722249 . _:b547722217 . _:b547722218 . _:b547722219 . . _:b547722212 . _:b547722213 . _:b547722248 . _:b547722214 . . _:b547722215 . _:b547722208 . _:b547722209 . _:b547722251 . _:b547722210 . _:b547722453 . _:b29675284 "Results from several cell culture studies have suggested that HBV can modulate monocyte activation resulting in induction of M2 associated cytokines and inhibition of M1 associated cytokines [20], [22], [47], [>>48<<], [49]. Here we demonstrate that HBV viral stock promoted M2-like macrophage activation in both human M1 and M2 polarized macrophages as examined by the induction of spindle/fibroblast shaped morphology (as opposed to round/oval shaped" . _:b547722211 . _:b547722204 . . _:b547722205 . _:b547722250 . _:b547722206 . _:b547722207 . _:b547722113 . _:b547722200 . _:b547722201 . _:b547722261 . _:b547722202 . _:b547722203 . _:b547722196 . _:b547722197 . _:b29675271 . _:b547722260 . . _:b547722198 . _:b547722199 . _:b547722192 . _:b547722193 . _:b547722263 . _:b547722194 . _:b547722470 . _:b547722195 . _:b547722188 . _:b547722366 . _:b547722189 . _:b547722262 . _:b547722190 . _:b547722191 . _:b547722577 . _:b547722184 . . _:b547722436 . _:b547722433 . _:b547722257 . _:b547722185 . _:b547722186 . _:b547722187 . _:b547722180 . _:b547722181 . _:b547722256 . _:b547722182 . _:b547722183 . _:b547722176 . _:b547722177 . _:b547722259 . . _:b547722178 . _:b547722179 . _:b547722258 . _:b547722269 . _:b547722610 . _:b547722268 . . _:b29675250 . _:b547722439 . _:b547722271 . _:b547722486 . _:b547722270 . . _:b547722265 . . _:b547722264 . _:b547722254 . _:b547722267 . _:b547722209 . _:b547722133 . _:b547722266 . . _:b29675205 "Chronic hepatitis B virus (HBV) infection results in liver fibrosis/cirrhosis and development of hepatocellular carcinoma (HCC) [>>1<<], [2]. Establishment of chronic HBV infection is inversely associated with patient's age with neonatal and infants most susceptible, while adults are mostly resistant to chronic infection [3], [4]." . _:b547722277 . _:b547722485 . _:b547722276 . _:b547722393 . _:b547722279 . _:b547722589 . _:b547722278 . _:b547722423 . _:b547722273 . . _:b547722272 . _:b547722124 . _:b547722275 . . _:b547722108 . _:b547722109 . _:b547722274 . _:b547722110 . _:b547722111 . _:b547722285 . _:b547722106 . _:b547722107 . _:b547722284 . _:b547722560 . _:b29675250 "Human liver progenitor cells containing hepatoblasts (Hep) and CD34+ hematopoietic stem cells (HSC) were isolated from 15\u201319 weeks old human fetal liver tissue (Advanced Bioscience Resources) essentially as described [56], [>>57<<], [58]. To separate progenitor liver cells from non-parenchymal cells (including HSC), the fetal liver cells were centrifuged at low speed three times (5 minutes, 18\u00D7g)." . _:b547722287 . . _:b547722286 . _:b547722281 . . _:b547722280 . . _:b547722283 . . _:b547722282 . _:b547722293 . . _:b547722441 . . _:b547722292 . _:b547722295 . . _:b547722294 . . _:b547722289 . . . _:b547722288 . _:b547722502 . . _:b547722291 . _:b547722440 . _:b547722290 . _:b547722125 . _:b547722301 . _:b547722265 . _:b547722300 . _:b547722303 . _:b29675272 "AMAC1 - alternative macrophage activation-associated CC chemokine-1, IL10, B7-H4) and down-regulation or no change of M1-like macrophage genes (TNF\u03B1, iNOS, IL12p40) in HBV infected patients compared to healthy controls (Figure S8, S9) [>>8<<], [9]. Furthermore, liver gene expression profile analysis showed M2-like macrophage gene expression profile (CD68high, CD163high, AMAC1high, iNOSlow, TNF\u03B1low) is associated with upregulation of tissue fibrosis (COL1A1high, TIMP1high) and" . _:b29675244 . . _:b547722302 . _:b547722297 . . _:b547722296 . _:b547722299 . _:b547722298 . _:b547722508 . _:b547722309 . _:b547722308 . _:b547722311 . _:b547722145 . _:b547722310 . _:b547722305 . . _:b547722304 . _:b29675211 . _:b547722307 . _:b547722306 . _:b547722132 . _:b547722317 . . _:b547722497 . . _:b547722316 . . _:b547722319 . _:b547722318 . _:b547722490 . _:b547722313 . . . . _:b547722312 . _:b547722588 . _:b547722315 . _:b547722203 . _:b547722314 . _:b547722325 . _:b547722324 . "PMC0" . . _:b547722327 . _:b29675276 . _:b547722326 . . _:b547722321 . _:b547722221 . _:b547722320 . _:b547722598 . _:b547722323 . . _:b547722322 . _:b547722333 . . _:b547722332 . _:b547722335 . _:b547722334 . _:b547722474 . _:b547722329 . . _:b547722328 . . _:b547722331 . . . _:b547722330 . _:b547722341 . _:b547722340 . _:b547722202 . . _:b547722343 . _:b29675308 "Several studies have reported that HBV virus/HBV-encoded proteins can directly promote M2-like activation [20], [>>21<<], [22]. We confirmed and extended those results by demonstrating that HBV promotes M2 macrophage polarization in human M1 and M2 macrophages." . . . _:b547722342 . _:b547722337 . . _:b547722455 . _:b547722336 . . _:b29675296 "Genetic modification of human liver cells for enhanced survival, repopulation and differentiation coupled with mouse Fas agonist (Jo2) [>>34<<] and/or the AFC8 murine liver damage system [18] could further enhance human liver repopulation." . _:b547722339 . _:b547722338 . . _:b547722349 . _:b547722190 . _:b29675261 . _:b547722348 . _:b29675247 . . _:b547722351 . _:b547722350 . _:b547722277 . _:b547722345 . . _:b29675259 . _:b547722344 . _:b547722347 . . _:b547722570 . _:b547722346 . _:b547722357 . . _:b547722356 . _:b547722525 . _:b547722295 . _:b29675285 . _:b547722359 . _:b547722358 . . _:b547722353 . _:b547722352 . _:b547722487 . . _:b547722355 . _:b29675216 . _:b547722354 . _:b547722446 . _:b547722365 . _:b547722309 . . _:b547722364 . _:b29675263 . . _:b547722548 . . _:b547722367 . _:b29675262 . _:b547722366 . . _:b547722361 . _:b547722426 . _:b547722360 . _:b547722363 . . _:b547722281 . . _:b547722362 . _:b547722641 . _:b547722373 . _:b547722230 . _:b547722180 . _:b547722444 . _:b547722372 . . _:b547722375 . . _:b547722374 . _:b547722369 . _:b547722327 . . _:b547722368 . . . _:b547722371 . _:b547722370 . _:b547722187 . _:b547722523 . _:b547722381 . _:b547722291 . _:b547722380 . _:b547722383 . . _:b547722251 . _:b547722382 . . . _:b547722377 . _:b547722567 . _:b547722376 . _:b547722379 . . _:b547722378 . _:b547722542 . _:b547722389 . _:b547722503 . _:b547722388 . _:b547722391 . _:b547722390 . _:b547722110 . . _:b547722385 . _:b547722654 . _:b547722384 . _:b547722387 . . _:b547722386 . . _:b29675292 . _:b29675278 . _:b547722397 . . _:b547722122 . _:b547722396 . _:b547722232 . _:b547722399 . _:b547722318 . . _:b547722398 . _:b547722126 . _:b547722393 . . _:b547722392 . _:b547722395 . . _:b547722394 . _:b547722405 . . _:b29675268 "Several studies have suggested liver specific T cell immune impairment in chronic HBV infection [>>5<<]. To characterize anti-HBV human T cell immune response in lymphoid and liver tissues, HLA-A2 donor derived-leukocytes from the spleen and lymph nodes or livers of mock-, HBV plus neutralizing antibody- or HBV-infected humanized animals" . _:b547722404 . _:b547722407 . _:b547722406 . . _:b547722401 . _:b547722416 . _:b547722400 . _:b547722403 . . _:b29675274 "expression profile analysis showed M2-like macrophage gene expression profile (CD68high, CD163high, AMAC1high, iNOSlow, TNF\u03B1low) is associated with upregulation of tissue fibrosis (COL1A1high, TIMP1high) and damage markers (HMGB1high [>>42<<], [43], [44], S100A9high [45], [46]) (Figure S9) [8], [9]." . _:b547722613 . _:b547722402 . _:b547722204 . _:b547722413 . _:b547722412 . . _:b547722415 . _:b547722414 . . . _:b547722409 . _:b547722408 . . _:b547722411 . _:b547722467 . _:b29675249 "Human liver progenitor cells containing hepatoblasts (Hep) and CD34+ hematopoietic stem cells (HSC) were isolated from 15\u201319 weeks old human fetal liver tissue (Advanced Bioscience Resources) essentially as described [>>56<<], [57], [58]. To separate progenitor liver cells from non-parenchymal cells (including HSC), the fetal liver cells were centrifuged at low speed three times (5 minutes, 18\u00D7g)." . _:b547722410 . _:b547722421 . _:b547722325 . _:b29675220 . _:b547722420 . . _:b547722423 . . _:b547722422 . _:b547722576 . _:b547722417 . . _:b547722416 . _:b547722419 . _:b547722418 . _:b547722555 . _:b547722429 . _:b547722428 . _:b547722431 . _:b547722484 . . _:b547722430 . _:b547722130 . _:b547722425 . . _:b547722424 . . . _:b547722427 . . _:b547722426 . . _:b547722437 . . _:b547722436 . . _:b547722349 . _:b547722439 . . _:b547722438 . _:b547722433 . . _:b547722432 . . _:b547722435 . . _:b547722650 . _:b547722672 . _:b547722670 . _:b547722434 . . _:b547722233 . _:b547722445 . . _:b547722389 . _:b547722444 . _:b547722119 . _:b547722447 . _:b547722387 . . _:b547722446 . _:b547722441 . _:b547722581 . _:b547722440 . _:b547722565 . _:b547722443 . _:b29675243 . . _:b547722194 . . _:b547722442 . _:b547722453 . _:b29675280 . _:b547722452 . . _:b547722455 . _:b547722511 . _:b547722454 . . _:b547722266 . _:b547722449 . _:b547722160 . _:b547722448 . . _:b547722245 . _:b547722451 . . . _:b547722450 . _:b29675273 . . _:b547722461 . _:b547722460 . _:b547722463 . _:b547722462 . _:b547722457 . _:b547722123 . _:b547722456 . . _:b547722586 . _:b547722459 . . _:b547722458 . _:b547722469 . . . _:b547722468 . _:b547722471 . _:b29675264 "results" . _:b547722404 . . . _:b547722470 . _:b547722430 . _:b547722465 . _:b547722464 . _:b547722405 . _:b547722467 . _:b29675288 "However, these mice have disadvantages including neonatal death, poor health and, most importantly, the lack of a human immune system [>>19<<]. To overcome these deficiencies, the A2/NSG/Fas-humanized mouse model enables inducible depletion of murine hepatocytes through the Fas apoptotic signaling pathway, resulting in elevated human liver repopulation in mice transplanted with" . _:b547722466 . _:b547722477 . . _:b29675245 . _:b547722476 . _:b547722505 . . _:b547722479 . _:b29675243 "To promote human liver cell repopulation, A2/NSG-hu HSC/Hep mice were treated with a murine specific anti-Fas agonistic antibody (Jo2) [32], [33], [>>34<<], [35], [36]." . _:b547722478 . _:b29675304 "tissue fibrosis and damage; with M1 polarized macrophages promoting anti-virus Th1 immune response and pathogen clearance, while M2 polarized macrophages impair Th1 immune response and promoting tissue remodeling [23], [24], [25], [>>26<<], [27]. M2 macrophages are critical innate immune cells involved in tissue remodeling/wound repair, secreting anti-inflammatory cytokines and redistributing micronutrients to sites of wound repair; however, during chronic infection," . _:b547722111 "11"^^ . _:b547722213 . _:b547722473 . _:b547722110 "11"^^ . _:b547722442 . _:b547722205 . . . _:b547722472 . . . _:b547722109 "12"^^ . _:b547722106 "16"^^ . _:b547722362 . _:b547722475 . _:b547722107 "16"^^ . . _:b547722108 "13"^^ . _:b547722517 . . _:b547722474 . . _:b547722485 . _:b547722640 . _:b547722484 . _:b547722270 . _:b547722624 . _:b547722487 . _:b29675221 . _:b547722486 . _:b547722118 "8"^^ . _:b547722481 . _:b547722119 "7"^^ . _:b29675223 . _:b547722480 . . _:b547722116 "8"^^ . . . _:b547722483 . _:b547722117 "8"^^ . _:b547722482 . . _:b547722114 "8"^^ . _:b29675210 "Though HBV-induced liver disease is predominately a chronic disease, requiring decades of chronic infection and liver inflammation [5], [6], [>>7<<], HBV infection occasionally results in accelerated liver disease and liver failure during acute infection [8], [9]." . _:b547722493 . _:b547722115 "8"^^ . _:b547722492 . _:b547722151 . _:b547722112 "10"^^ . _:b547722495 . _:b547722113 "9"^^ . . _:b547722126 "6"^^ . _:b547722494 . . _:b547722582 . _:b547722127 "6"^^ . _:b547722489 . . _:b29675215 . _:b547722124 "7"^^ . _:b547722488 . _:b547722491 . _:b547722125 "7"^^ . _:b547722192 . _:b547722490 . _:b547722122 "7"^^ . _:b547722558 . . _:b547722123 "7"^^ . . _:b547722120 "7"^^ . _:b547722449 . _:b29675264 . . _:b547722121 "7"^^ . _:b547722134 "6"^^ . _:b547722135 "6"^^ . . _:b547722132 "6"^^ . _:b29675296 . . _:b29675219 . _:b29675286 . _:b29675206 . _:b547722133 "6"^^ . . _:b547722130 "6"^^ . . _:b547722131 "6"^^ . _:b547722128 "6"^^ . . . _:b547722129 "6"^^ . _:b547722207 . _:b29675295 . . . _:b547722189 . _:b547722141 "5"^^ . _:b547722304 . . _:b547722140 "5"^^ . _:b547722143 "5"^^ . . _:b29675238 "They support efficient development of a functional human immune system after injecting CD34+ human hematopoietic stem cells (HSC) into the liver of newborn mice [>>30<<], [31]. Furthermore, the A2/NSG mouse carries the human HLA-A2 transgene, which enhances development of human MHC-restrict T lymphocytes [30]. To promote human liver cell repopulation, A2/NSG-hu HSC/Hep mice were treated with a murine" . . _:b547722142 "5"^^ . _:b547722138 "6"^^ . _:b547722136 "6"^^ . _:b547722139 "5"^^ . _:b547722137 "6"^^ . _:b547722398 . . _:b547722149 "5"^^ . _:b547722148 "5"^^ . . _:b547722151 "5"^^ . . _:b547722150 "5"^^ . _:b547722145 "5"^^ . _:b547722144 "5"^^ . _:b547722335 . _:b547722571 . _:b547722147 "5"^^ . _:b547722526 . _:b547722146 "5"^^ . _:b547722157 "5"^^ . _:b547722333 . _:b547722156 "5"^^ . _:b547722159 "5"^^ . _:b29675291 . _:b547722158 "5"^^ . _:b547722533 . _:b547722153 "5"^^ . _:b547722152 "5"^^ . . _:b29675294 "Additionally, the A2/NSG background permits highly efficient engraftment and development of human xenografts including human hematopoietic stem cells compared to current immunodeficient mouse models [>>52<<]. Lower levels of human hepatocytes were detected in A2/NSG/Fas-humanized mice in comparison to the uPA or FAH mice transplanted with adult human hepatocytes. However, it should be noted that fetal liver cell repopulation is also low in" . _:b547722155 "5"^^ . _:b547722210 . _:b547722154 "5"^^ . . _:b547722165 "5"^^ . _:b547722164 "5"^^ . . _:b547722488 . _:b547722167 "4"^^ . _:b547722639 . _:b547722458 . _:b547722166 "4"^^ . . _:b547722150 . _:b547722161 "5"^^ . _:b29675246 "materials and methods" . _:b547722160 "5"^^ . _:b29675284 . _:b547722163 "5"^^ . . _:b547722162 "5"^^ . . _:b547722173 "4"^^ . . _:b547722172 "4"^^ . . _:b547722175 "4"^^ . _:b547722459 . _:b547722174 "4"^^ . _:b547722397 . _:b547722169 "4"^^ . . _:b547722415 . _:b547722168 "4"^^ . . _:b547722171 "4"^^ . _:b29675241 . . _:b547722170 "4"^^ . . _:b547722653 . _:b547722510 . _:b547722181 "4"^^ . _:b547722271 . _:b547722186 . _:b29675290 . _:b547722180 "4"^^ . _:b29675300 . _:b547722183 "4"^^ . _:b547722409 . _:b29675266 . _:b547722183 . . _:b547722182 "4"^^ . _:b547722673 . _:b547722177 "4"^^ . _:b547722176 "4"^^ . _:b29675297 . _:b547722491 . _:b547722179 "4"^^ . . _:b29675289 "mouse model enables inducible depletion of murine hepatocytes through the Fas apoptotic signaling pathway, resulting in elevated human liver repopulation in mice transplanted with human liver progenitor and hematopoietic stem cells [>>32<<], [33], [34], [36], [51]. Additionally, the A2/NSG background permits highly efficient engraftment and development of human xenografts including human hematopoietic stem cells compared to current immunodeficient mouse models [52]." . _:b547722178 "4"^^ . _:b547722376 . _:b547722189 "4"^^ . _:b547722109 . _:b547722188 "4"^^ . _:b547722399 . _:b547722544 . _:b547722191 "4"^^ . _:b547722190 "4"^^ . _:b547722596 . _:b547722185 "4"^^ . _:b547722352 . _:b547722184 "4"^^ . _:b547722187 "4"^^ . _:b547722248 . _:b547722186 "4"^^ . _:b547722296 . _:b547722197 "4"^^ . _:b29675242 . _:b29675287 . _:b547722196 "4"^^ . _:b547722199 "4"^^ . _:b547722198 "4"^^ . _:b547722382 . . _:b547722193 "4"^^ . _:b29675246 . _:b547722628 . _:b547722192 "4"^^ . _:b547722195 "4"^^ . _:b547722614 . . _:b547722194 "4"^^ . _:b547722205 "3"^^ . . . _:b547722204 "3"^^ . _:b547722600 . _:b29675204 . _:b547722313 . _:b547722207 "3"^^ . _:b29675205 . . _:b547722206 "3"^^ . _:b29675206 . _:b547722201 "3"^^ . _:b29675207 . _:b547722634 . _:b547722200 "4"^^ . _:b29675204 . _:b547722203 "3"^^ . . _:b547722288 . _:b547722202 "3"^^ . _:b29675207 "Establishment of chronic HBV infection is inversely associated with patient's age with neonatal and infants most susceptible, while adults are mostly resistant to chronic infection [3], [>>4<<]. Chronic HBV infection is associated with impaired immune responses to viral antigens and chronic inflammation in the liver, leading to progressive liver diseases. Though HBV-induced liver disease is predominately a chronic disease," . _:b547722394 . . _:b547722213 "3"^^ . _:b547722585 . _:b547722212 "3"^^ . _:b29675212 . _:b547722215 "3"^^ . _:b29675213 . . . _:b547722214 "3"^^ . _:b547722303 . _:b29675214 . _:b547722209 "3"^^ . _:b29675215 . _:b547722208 "3"^^ . _:b29675208 . _:b29675309 "Several studies have reported that HBV virus/HBV-encoded proteins can directly promote M2-like activation [20], [21], [>>22<<]. We confirmed and extended those results by demonstrating that HBV promotes M2 macrophage polarization in human M1 and M2 macrophages." . _:b547722211 "3"^^ . _:b29675209 . _:b547722210 "3"^^ . . _:b29675210 . . _:b547722221 "3"^^ . _:b547722227 . _:b29675255 "RNA was isolated from tissue following manufacturer's recommended procedures and qPCR was performed using species-specific published [18], [>>61<<] or NCBI primer blast generated primers and the SYBR Green method, following manufacturer's recommended procedures (Thermo Scientific)." . _:b29675211 . _:b547722220 "3"^^ . _:b29675220 . . . _:b547722223 "3"^^ . . _:b29675221 . _:b547722222 "3"^^ . _:b29675222 . _:b547722217 "3"^^ . _:b29675223 . _:b547722216 "3"^^ . _:b29675216 . _:b547722332 . _:b547722219 "3"^^ . _:b547722609 . _:b29675217 . _:b547722218 "3"^^ . _:b29675218 . _:b547722229 "3"^^ . _:b29675219 . . _:b547722228 "3"^^ . . _:b29675228 . _:b547722656 . _:b547722532 . . _:b547722231 "3"^^ . _:b547722545 . _:b29675229 . _:b547722257 . _:b547722230 "3"^^ . . _:b29675230 . . _:b547722225 "3"^^ . _:b29675231 . . _:b547722224 "3"^^ . _:b29675224 . . _:b547722227 "3"^^ . _:b29675225 . _:b547722543 . _:b547722226 "3"^^ . . _:b29675226 . _:b547722572 . _:b547722237 "3"^^ . . _:b29675227 . _:b547722236 "3"^^ . _:b29675236 . _:b547722184 . _:b547722138 . _:b547722239 "3"^^ . _:b29675237 . _:b547722388 . _:b547722365 . _:b547722238 "3"^^ . _:b29675238 . _:b547722606 . _:b547722233 "3"^^ . _:b29675239 . . _:b547722232 "3"^^ . _:b29675232 . . _:b547722235 "3"^^ . _:b29675233 . . . . _:b547722524 . _:b547722234 "3"^^ . _:b29675234 . _:b29675235 . . _:b29675244 . _:b547722608 . . _:b29675245 . . _:b29675236 . _:b29675246 . . _:b29675247 . _:b547722599 . . _:b547722109 . _:b547722128 . _:b29675240 . _:b547722108 . _:b29675241 . . _:b547722111 . _:b29675242 . _:b547722110 . _:b29675243 . _:b29675218 "The Alb-uPA/SCID humanized mouse with high human adult hepatocyte repopulation can be infected with HCV/HBV [>>14<<]. Additionally, the fumarylacetoacetate hydrolase (Fah)-Rag2-\u03B3C-null mice also allow human hepatocytes engraftment and HCV infection [15], [16], [17]. However, these human-murine chimeric liver models lack a functional human immune system," . _:b29675252 . . _:b29675253 . _:b547722107 . _:b29675254 . _:b547722573 . . _:b547722106 . . _:b29675255 . _:b547722117 . . _:b29675248 . _:b547722116 . _:b29675249 . _:b547722431 . _:b547722119 . _:b29675250 . _:b547722140 . _:b547722118 . _:b29675251 . _:b547722113 . _:b29675260 . _:b547722112 . _:b29675261 . . _:b547722539 . _:b547722115 . . _:b29675262 . _:b547722108 . _:b547722114 . . _:b29675263 . _:b547722546 . _:b29675205 . _:b547722125 . _:b29675256 . . _:b547722124 . _:b29675257 . . _:b547722127 . _:b547722324 . _:b29675258 . _:b547722126 . . _:b547722280 . _:b29675259 . _:b547722234 . . _:b547722121 . _:b547722516 . _:b547722120 . . _:b547722123 . . _:b547722122 . _:b547722133 . . _:b547722132 . . . _:b547722135 . . _:b547722134 . _:b29675307 "Several studies have reported that HBV virus/HBV-encoded proteins can directly promote M2-like activation [>>20<<], [21], [22]." . . . . _:b547722129 . _:b547722128 . _:b547722131 . _:b547722604 . _:b547722130 . _:b547722141 . . _:b547722140 . _:b547722341 . _:b547722143 . _:b547722622 . _:b547722142 . _:b547722137 . . . _:b547722136 . _:b547722139 . _:b547722138 . _:b547722177 . _:b547722149 . _:b547722348 . _:b547722148 . _:b547722151 . _:b547722150 . . _:b547722145 . _:b547722144 . _:b29675247 "Liver gene expression profile analyses in patients were obtained from a dataset in Gene Expression Omnibus (GEO)/NCBI database; the reports followed NIH research ethics guidelines [>>8<<], [9]." . _:b547722564 . . _:b547722147 . _:b547722146 . . _:b547722157 . _:b547722538 . _:b547722156 . _:b547722645 . _:b547722159 . _:b547722617 . _:b547722158 . _:b547722153 . . . . _:b547722152 . . _:b547722155 . _:b547722164 . _:b547722509 . _:b547722154 . _:b547722637 . _:b547722165 . _:b547722164 . . _:b547722167 . _:b547722166 . _:b547722161 . _:b547722554 . _:b547722160 . _:b547722515 . _:b547722163 . _:b547722162 . . _:b547722173 . . _:b29675237 "mouse model by injecting human liver progenitor cells (Hep) and CD34+ human hematopoietic stem cells (HSC) directly into the liver of newborn A2/NSG (HLA-A2 transgenic NOD scid IL2 receptor gamma chain knockout mice [28], [29], [>>30<<]). The A2/NSG mouse lacks NK cells and T/B-lymphocytes. They support efficient development of a functional human immune system after injecting CD34+ human hematopoietic stem cells (HSC) into the liver of newborn mice [30], [31]." . . _:b547722172 . _:b547722175 . _:b547722174 . _:b547722169 . _:b547722664 . _:b547722168 . _:b29675238 . _:b547722171 . _:b547722258 . _:b547722170 . _:b547722181 . . . _:b29675237 . _:b547722180 . _:b547722574 . . _:b29675206 "Chronic hepatitis B virus (HBV) infection results in liver fibrosis/cirrhosis and development of hepatocellular carcinoma (HCC) [1], [>>2<<]. Establishment of chronic HBV infection is inversely associated with patient's age with neonatal and infants most susceptible, while adults are mostly resistant to chronic infection [3], [4]." . _:b547722183 . _:b547722182 . _:b29675240 . _:b547722177 . _:b547722176 . _:b547722179 . _:b547722165 . . _:b547722178 . . _:b29675283 "Results from several cell culture studies have suggested that HBV can modulate monocyte activation resulting in induction of M2 associated cytokines and inhibition of M1 associated cytokines [20], [22], [>>47<<], [48], [49]." . _:b547722189 . _:b547722188 . _:b29675265 "We confirmed the specie-specificity of Jo2 antibody [>>32<<] by incubating human liver cell line (HepG2) with Jo2 antibody." . _:b547722191 . _:b547722190 . _:b547722354 . _:b547722185 . _:b547722184 . . _:b547722343 . . _:b547722429 . _:b547722187 . . . _:b547722186 . _:b547722197 . _:b547722196 . . _:b547722451 . . _:b547722199 . _:b547722479 . _:b547722198 . _:b547722193 . _:b547722192 . _:b547722212 . _:b547722195 . _:b547722166 . _:b29675226 "Additionally, HCV infection induces liver inflammation and fibrosis, correlated with activation of human hepatic stellate cells and expression of human fibrogenic genes [>>18<<]." . _:b547722194 . _:b547722536 . _:b547722205 . _:b547722204 . . _:b547722207 . _:b547722206 . . _:b547722201 . _:b547722200 . . _:b547722203 . _:b547722386 . . _:b547722202 . . _:b547722225 . . _:b547722213 . _:b547722212 . . _:b547722238 . _:b29675278 "M2-like macrophage gene expression profile (CD68high, CD163high, AMAC1high, iNOSlow, TNF\u03B1low) is associated with upregulation of tissue fibrosis (COL1A1high, TIMP1high) and damage markers (HMGB1high [42], [43], [44], S100A9high [45], [>>46<<]) (Figure S9) [8], [9]." . _:b547722215 . _:b547722214 . _:b547722209 . _:b547722553 . . _:b547722208 . _:b547722211 . _:b547722210 . _:b547722580 . _:b547722221 . _:b547722668 . . _:b547722220 . _:b547722223 . . _:b547722222 . _:b547722217 . _:b547722216 . . _:b547722219 . _:b547722218 . . _:b547722229 . _:b547722500 . _:b547722228 . . . _:b547722231 . _:b547722230 . _:b547722225 . _:b547722216 . . _:b547722297 . _:b547722120 . . _:b547722224 . . _:b29675230 "pathogen clearance, chronic inflammation and associated liver pathology; with M1 polarized macrophages promoting pathogen clearance, and M2-like polarized macrophages impairing host immunity and promoting tissue fibrosis/remodeling [>>23<<], [24], [25], [26], [27]." . _:b29675301 . . _:b547722227 . _:b547722121 . _:b547722226 . _:b29675240 "Furthermore, the A2/NSG mouse carries the human HLA-A2 transgene, which enhances development of human MHC-restrict T lymphocytes [>>30<<]. To promote human liver cell repopulation, A2/NSG-hu HSC/Hep mice were treated with a murine specific anti-Fas agonistic antibody (Jo2) [32], [33], [34], [35], [36]. The A2/NSG-hu HSC/Hep mouse model enabled human liver and immune system" . _:b547722237 . . _:b547722236 . . _:b29675257 "Immunoreactivity was determined by incubation with DAB substrate (Pierce) or Vulcan red (Dako), and counterstained with hematoxilin [56], [>>58<<]. Liver histological activity was determined using the knodell score, which examines liver necrosis, degeneration, inflammation and fibrosis [62], [63]." . . _:b547722239 . . . _:b547722238 . _:b547722233 . _:b547722232 . . _:b547722235 . . . _:b547722234 . . _:b547722197 . _:b547722469 . . . _:b547722334 . _:b29675249 . . _:b547722148 . _:b29675305 "fibrosis and damage; with M1 polarized macrophages promoting anti-virus Th1 immune response and pathogen clearance, while M2 polarized macrophages impair Th1 immune response and promoting tissue remodeling [23], [24], [25], [26], [>>27<<]. M2 macrophages are critical innate immune cells involved in tissue remodeling/wound repair, secreting anti-inflammatory cytokines and redistributing micronutrients to sites of wound repair; however, during chronic infection, M2-like" . _:b547722638 . _:b547722633 . _:b29675256 . . . _:b547722300 . _:b547722521 . _:b547722114 . . . _:b547722559 . _:b29675270 "suppressor cells expressing high levels of IL10, co-inhibitory molecules (B7-H4), while depleting L-arginine and down-regulating IL12, TNF\u03B1 and co-stimulatory molecules (CD86); all factors critical for Th1 anti-viral immune response [>>41<<]. Analysis of liver inflammation in HBV infected humanized mice showed human M2 macrophages co-localized with human T cells (Figure 7C, Table S2)." . . . _:b29675286 "discussion" . . _:b547722176 . _:b547722381 . . _:b547722496 . . . _:b547722226 . . _:b547722252 . _:b29675239 "They support efficient development of a functional human immune system after injecting CD34+ human hematopoietic stem cells (HSC) into the liver of newborn mice [30], [>>31<<]. Furthermore, the A2/NSG mouse carries the human HLA-A2 transgene, which enhances development of human MHC-restrict T lymphocytes [30]. To promote human liver cell repopulation, A2/NSG-hu HSC/Hep mice were treated with a murine specific" . _:b547722632 . _:b547722307 . _:b547722454 . _:b547722465 . _:b547722129 . _:b29675259 "Liver histological activity was determined using the knodell score, which examines liver necrosis, degeneration, inflammation and fibrosis [62], [>>63<<]." . _:b547722346 . . _:b547722563 . _:b547722413 . _:b547722360 . _:b547722476 . . . . . . _:b547722171 . . _:b547722592 . . . . _:b547722135 . . . _:b547722493 . _:b547722535 . . _:b547722314 . . _:b29675220 "Additionally, the fumarylacetoacetate hydrolase (Fah)-Rag2-\u03B3C-null mice also allow human hepatocytes engraftment and HCV infection [15], [>>16<<], [17]. However, these human-murine chimeric liver models lack a functional human immune system, thus it is not possible to study host immune response and hepatitis virus-induced immunopathology [14], [17]. To overcome the limitations" . _:b547722260 . _:b547722244 . . _:b547722355 . _:b547722447 . . _:b547722168 . . . _:b547722342 . _:b29675235 . . _:b29675258 "Liver histological activity was determined using the knodell score, which examines liver necrosis, degeneration, inflammation and fibrosis [>>62<<], [63]." . . _:b547722330 . . _:b547722267 . _:b547722462 . _:b547722478 . . _:b547722677 . _:b547722461 . _:b547722358 . _:b547722374 . _:b547722245 "3"^^ . . _:b547722244 "3"^^ . _:b547722410 . _:b547722247 "3"^^ . . _:b547722246 "3"^^ . _:b547722308 . _:b547722569 . _:b547722241 "3"^^ . _:b547722211 . _:b547722240 "3"^^ . . _:b547722243 "3"^^ . _:b547722181 . _:b547722242 "3"^^ . _:b547722253 "3"^^ . . _:b547722252 "3"^^ . _:b547722255 "3"^^ . _:b547722463 . _:b29675227 "Several reports suggest that HBV promotes macrophage activation and M2 polarization [>>20<<], [21], [22]." . _:b547722254 "3"^^ . _:b547722249 "3"^^ . _:b547722253 . . _:b547722248 "3"^^ . _:b547722251 "3"^^ . . _:b547722250 "3"^^ . _:b29675222 "However, these human-murine chimeric liver models lack a functional human immune system, thus it is not possible to study host immune response and hepatitis virus-induced immunopathology [>>14<<], [17]. To overcome the limitations associated with current chimeric human-murine liver mouse models, we have recently developed a humanized mouse model with both human immune system and liver cells (AFC8-hu HSC/Hep mice) [18], [19]." . . _:b547722261 "3"^^ . . _:b29675223 "However, these human-murine chimeric liver models lack a functional human immune system, thus it is not possible to study host immune response and hepatitis virus-induced immunopathology [14], [>>17<<]. To overcome the limitations associated with current chimeric human-murine liver mouse models, we have recently developed a humanized mouse model with both human immune system and liver cells (AFC8-hu HSC/Hep mice) [18], [19]. AFC8-hu" . _:b547722260 "3"^^ . . _:b547722287 . _:b547722263 "3"^^ . _:b547722320 . _:b547722262 "3"^^ . _:b547722257 "3"^^ . _:b547722256 "3"^^ . _:b547722259 "3"^^ . _:b547722258 "3"^^ . _:b547722269 "3"^^ . . _:b547722268 "3"^^ . _:b547722271 "3"^^ . . _:b547722270 "3"^^ . . . _:b547722265 "3"^^ . . _:b547722264 "3"^^ . _:b547722267 "3"^^ . _:b547722266 "3"^^ . _:b547722277 "3"^^ . _:b547722276 "3"^^ . . _:b29675248 . _:b547722279 "3"^^ . _:b547722278 "3"^^ . _:b547722273 "3"^^ . _:b547722306 . _:b29675253 . _:b547722272 "3"^^ . . _:b547722134 . _:b547722275 "3"^^ . . _:b547722274 "3"^^ . _:b547722338 . _:b29675274 . _:b547722285 "3"^^ . _:b547722621 . _:b547722290 . _:b547722284 "3"^^ . . _:b547722139 . _:b547722287 "3"^^ . . . _:b547722286 "3"^^ . _:b547722281 "3"^^ . . _:b547722280 "3"^^ . _:b547722644 . . _:b547722283 "3"^^ . _:b547722345 . _:b547722282 "3"^^ . . _:b547722293 "3"^^ . _:b547722406 . . _:b547722292 "3"^^ . . _:b547722295 "3"^^ . _:b547722294 "3"^^ . . _:b547722289 "3"^^ . . _:b547722288 "3"^^ . . _:b29675208 "Though HBV-induced liver disease is predominately a chronic disease, requiring decades of chronic infection and liver inflammation [>>5<<], [6], [7], HBV infection occasionally results in accelerated liver disease and liver failure during acute infection [8], [9]." . _:b547722422 . _:b547722291 "3"^^ . . _:b547722290 "3"^^ . _:b547722301 "3"^^ . . _:b547722379 . . _:b547722300 "3"^^ . . _:b547722275 . _:b547722303 "3"^^ . _:b547722193 . _:b547722302 "3"^^ . _:b547722648 . _:b547722297 "3"^^ . _:b547722222 . _:b547722296 "3"^^ . _:b29675305 . _:b547722299 "3"^^ . . _:b547722667 . _:b547722298 "3"^^ . _:b547722309 "3"^^ . _:b29675307 . _:b547722642 . _:b547722308 "3"^^ . _:b547722317 . _:b547722311 "2"^^ . _:b547722489 . _:b547722310 "2"^^ . _:b547722178 . _:b547722305 "3"^^ . _:b547722304 "3"^^ . _:b29675213 . _:b547722307 "3"^^ . . . _:b547722306 "3"^^ . . _:b29675275 . _:b547722317 "2"^^ . _:b547722391 . _:b547722316 "2"^^ . _:b547722629 . _:b547722319 "2"^^ . . _:b547722318 "2"^^ . _:b547722188 . _:b29675292 "inducible depletion of murine hepatocytes through the Fas apoptotic signaling pathway, resulting in elevated human liver repopulation in mice transplanted with human liver progenitor and hematopoietic stem cells [32], [33], [34], [>>36<<], [51]. Additionally, the A2/NSG background permits highly efficient engraftment and development of human xenografts including human hematopoietic stem cells compared to current immunodeficient mouse models [52]." . _:b547722537 . _:b29675282 . _:b547722313 "2"^^ . _:b547722438 . _:b547722159 . _:b29675216 "Delineation of the mechanisms by which HBV evades host immunity to establish chronic infection and promote liver disease is hampered by the lack of robust animal models [11], [12], [>>13<<]." . _:b547722312 "2"^^ . . _:b547722471 . _:b547722315 "2"^^ . _:b29675212 . _:b547722314 "2"^^ . _:b547722325 "2"^^ . _:b547722324 "2"^^ . _:b547722327 "2"^^ . _:b547722326 "2"^^ . _:b547722321 "2"^^ . _:b547722320 "2"^^ . _:b547722289 . _:b547722477 . _:b547722323 "2"^^ . "10.1371%2Fjournal.ppat.1004032" . _:b29675303 . _:b547722322 "2"^^ . _:b547722333 "2"^^ . _:b547722332 "2"^^ . _:b547722143 . _:b547722335 "2"^^ . . . _:b547722198 . _:b547722334 "2"^^ . _:b547722561 . _:b547722403 . _:b29675287 "Immunodeficient mice expressing the uPA transgene in the liver of SCID mice or with mutant Fah genes allow transplanted human adult hepatocytes to have a growth advantage and efficiently repopulate the liver [>>50<<]. However, these mice have disadvantages including neonatal death, poor health and, most importantly, the lack of a human immune system [19]." . _:b547722329 "2"^^ . _:b547722361 . _:b547722328 "2"^^ . _:b547722315 . . _:b547722331 "2"^^ . _:b547722456 . _:b547722330 "2"^^ . _:b547722219 . _:b547722341 "2"^^ . _:b547722340 "2"^^ . _:b547722343 "2"^^ . . _:b547722342 "2"^^ . . _:b547722337 "2"^^ . _:b547722336 "2"^^ . . . . _:b547722339 "2"^^ . _:b547722338 "2"^^ . _:b547722349 "2"^^ . _:b547722331 . . _:b547722348 "2"^^ . _:b547722285 . _:b547722351 "2"^^ . . . _:b29675262 "17 different liver specimens approximately evenly taken from the 4 livers) and match healthy control liver donors (n\u200A=\u200A10); acute HBV- induced liver failure patients were previously healthy and had no signs of chronic liver disease [>>8<<]. Microarray dataset (GEO accession: GSE49656; [8]) was analyzed using GEOR (NCBI Software)." . _:b547722350 "2"^^ . . _:b547722345 "2"^^ . _:b547722344 "2"^^ . _:b29675228 "Several reports suggest that HBV promotes macrophage activation and M2 polarization [20], [>>21<<], [22]. Macrophages play a critical role in modulating pathogen clearance, chronic inflammation and associated liver pathology; with M1 polarized macrophages promoting pathogen clearance, and M2-like polarized macrophages impairing host" . _:b547722357 . _:b547722347 "2"^^ . . _:b547722346 "2"^^ . . _:b547722357 "2"^^ . _:b29675260 "All patients were diagnosed according to our previously described criteria [>>64<<], [65] and had not received any antiviral therapies or immunosuppressive drugs within six months before sampling." . _:b547722356 "2"^^ . _:b547722550 . _:b547722359 "2"^^ . _:b547722358 "2"^^ . _:b29675289 . _:b547722292 . _:b547722353 "2"^^ . . _:b547722352 "2"^^ . . _:b547722355 "2"^^ . _:b29675281 . _:b547722658 . _:b547722354 "2"^^ . _:b29675251 "Human liver progenitor cells containing hepatoblasts (Hep) and CD34+ hematopoietic stem cells (HSC) were isolated from 15\u201319 weeks old human fetal liver tissue (Advanced Bioscience Resources) essentially as described [56], [57], [>>58<<]. To separate progenitor liver cells from non-parenchymal cells (including HSC), the fetal liver cells were centrifuged at low speed three times (5 minutes, 18\u00D7g)." . . _:b547722365 "2"^^ . . _:b547722501 . _:b547722556 . _:b29675291 "enables inducible depletion of murine hepatocytes through the Fas apoptotic signaling pathway, resulting in elevated human liver repopulation in mice transplanted with human liver progenitor and hematopoietic stem cells [32], [33], [>>34<<], [36], [51]. Additionally, the A2/NSG background permits highly efficient engraftment and development of human xenografts including human hematopoietic stem cells compared to current immunodeficient mouse models [52]." . _:b547722364 "2"^^ . _:b547722367 "2"^^ . _:b547722529 . _:b29675285 "Results from several cell culture studies have suggested that HBV can modulate monocyte activation resulting in induction of M2 associated cytokines and inhibition of M1 associated cytokines [20], [22], [47], [48], [>>49<<]. Here we demonstrate that HBV viral stock promoted M2-like macrophage activation in both human M1 and M2 polarized macrophages as examined by the induction of spindle/fibroblast shaped morphology (as opposed to round/oval shaped" . _:b547722298 . _:b547722366 "2"^^ . _:b29675308 . _:b547722450 . _:b547722361 "2"^^ . _:b547722329 . _:b547722360 "2"^^ . _:b29675293 "depletion of murine hepatocytes through the Fas apoptotic signaling pathway, resulting in elevated human liver repopulation in mice transplanted with human liver progenitor and hematopoietic stem cells [32], [33], [34], [36], [>>51<<]. Additionally, the A2/NSG background permits highly efficient engraftment and development of human xenografts including human hematopoietic stem cells compared to current immunodeficient mouse models [52]." . . _:b547722363 "2"^^ . _:b547722362 "2"^^ . _:b547722373 "2"^^ . _:b547722154 . _:b29675309 . _:b547722372 "2"^^ . . _:b547722375 "2"^^ . . _:b547722374 "2"^^ . _:b547722282 . _:b547722369 "2"^^ . _:b547722283 . _:b29675229 "Several reports suggest that HBV promotes macrophage activation and M2 polarization [20], [21], [>>22<<]. Macrophages play a critical role in modulating pathogen clearance, chronic inflammation and associated liver pathology; with M1 polarized macrophages promoting pathogen clearance, and M2-like polarized macrophages impairing host immunity" . _:b547722368 "2"^^ . _:b547722371 "2"^^ . _:b29675234 . . _:b547722370 "2"^^ . . _:b547722326 . _:b547722381 "2"^^ . _:b547722380 "2"^^ . _:b547722328 . _:b547722383 "2"^^ . _:b29675257 . _:b547722382 "2"^^ . _:b29675265 . _:b547722284 . _:b547722377 "2"^^ . _:b547722562 . _:b29675217 "To overcome host species restriction barrier for in vivo infection and disease modeling, several human-murine chimeric liver models have been developed [>>14<<]. The Alb-uPA/SCID humanized mouse with high human adult hepatocyte repopulation can be infected with HCV/HBV [14]. Additionally, the fumarylacetoacetate hydrolase (Fah)-Rag2-\u03B3C-null mice also allow human hepatocytes engraftment and HCV" . _:b547722195 . _:b547722376 "2"^^ . _:b547722443 . _:b547722418 . _:b547722379 "2"^^ . . . _:b547722378 "2"^^ . _:b547722619 . _:b547722389 "2"^^ . _:b547722466 . _:b547722666 . _:b547722388 "2"^^ . . _:b547722391 "2"^^ . _:b547722390 "2"^^ . . _:b547722385 "2"^^ . _:b29675251 . . _:b547722384 "2"^^ . _:b547722387 "2"^^ . _:b547722386 "2"^^ . . . _:b547722397 "2"^^ . _:b547722173 . _:b547722396 "2"^^ . . _:b547722399 "2"^^ . . . . _:b547722398 "2"^^ . _:b547722534 . _:b29675214 "Delineation of the mechanisms by which HBV evades host immunity to establish chronic infection and promote liver disease is hampered by the lack of robust animal models [>>11<<], [12], [13]." . _:b29675302 . _:b547722393 "2"^^ . . _:b547722392 "2"^^ . . _:b547722395 "2"^^ . _:b547722394 "2"^^ . _:b547722405 "2"^^ . _:b547722179 . _:b547722404 "2"^^ . _:b547722615 . . _:b547722407 "2"^^ . _:b547722406 "2"^^ . _:b547722299 . _:b547722375 . _:b547722401 "2"^^ . . . _:b547722400 "2"^^ . . _:b547722199 . _:b547722390 . _:b547722403 "2"^^ . . _:b547722482 . _:b547722402 "2"^^ . _:b547722157 . _:b547722117 . _:b547722413 "2"^^ . _:b547722412 "2"^^ . . _:b29675260 . _:b547722595 . . _:b547722415 "2"^^ . _:b29675235 "a humanized mouse model by injecting human liver progenitor cells (Hep) and CD34+ human hematopoietic stem cells (HSC) directly into the liver of newborn A2/NSG (HLA-A2 transgenic NOD scid IL2 receptor gamma chain knockout mice [>>28<<], [29], [30]). The A2/NSG mouse lacks NK cells and T/B-lymphocytes. They support efficient development of a functional human immune system after injecting CD34+ human hematopoietic stem cells (HSC) into the liver of newborn mice [30], [31]." . _:b29675293 . _:b547722414 "2"^^ . . _:b547722409 "2"^^ . _:b547722408 "2"^^ . . _:b547722411 "2"^^ . _:b547722410 "2"^^ . . _:b547722421 "2"^^ . _:b29675267 . . _:b29675227 . _:b547722659 . _:b547722420 "2"^^ . _:b547722423 "2"^^ . _:b547722344 . _:b29675230 . . _:b29675269 "clearance, chronic inflammation and associated tissue pathology; with M1-like macrophages promoting pathogen clearance, and M2-like macrophages impairing Th1 immune response and promoting tissue fibrosis/remodeling/wound healing [>>40<<]. Immunohistochemical analysis of HBV associated liver inflammation in humanized mice showed high levels of human macrophages with predominately \u201CM2-like\u201D phenotype (hCD68high, hCD14high, hCD16low/medium, hCD163high, hCD206high," . _:b547722422 "2"^^ . _:b547722214 . _:b547722417 "2"^^ . _:b547722607 . _:b547722416 "2"^^ . _:b547722419 "2"^^ . _:b547722158 . _:b547722418 "2"^^ . _:b547722429 "2"^^ . _:b547722428 "2"^^ . _:b547722431 "2"^^ . _:b547722370 . . _:b547722430 "2"^^ . _:b547722528 . . _:b547722425 "2"^^ . _:b29675232 "clearance, chronic inflammation and associated liver pathology; with M1 polarized macrophages promoting pathogen clearance, and M2-like polarized macrophages impairing host immunity and promoting tissue fibrosis/remodeling [23], [24], [>>25<<], [26], [27]." . _:b547722424 "2"^^ . . _:b547722310 . _:b547722427 "2"^^ . . . _:b547722161 . _:b547722426 "2"^^ . _:b547722236 . _:b547722367 . _:b547722437 "2"^^ . _:b547722269 . _:b547722436 "2"^^ . _:b547722439 "2"^^ . _:b547722438 "2"^^ . . _:b547722268 . _:b547722433 "2"^^ . . _:b547722671 . . _:b547722432 "2"^^ . . _:b547722435 "2"^^ . . . . _:b547722434 "2"^^ . . _:b547722445 "2"^^ . _:b547722657 . _:b29675297 "Genetic modification of human liver cells for enhanced survival, repopulation and differentiation coupled with mouse Fas agonist (Jo2) [34] and/or the AFC8 murine liver damage system [>>18<<] could further enhance human liver repopulation." . _:b547722444 "2"^^ . . . _:b547722447 "2"^^ . . _:b547722446 "2"^^ . _:b547722527 . _:b547722441 "2"^^ . _:b547722440 "2"^^ . _:b547722301 . _:b547722443 "2"^^ . _:b547722182 . _:b547722442 "2"^^ . _:b547722506 . . _:b547722453 "2"^^ . _:b547722620 . . _:b547722452 "2"^^ . . . _:b29675207 . _:b547722551 . _:b547722455 "2"^^ . _:b29675263 "Microarray dataset (GEO accession: GSE49656; [>>8<<]) was analyzed using GEOR (NCBI Software)." . _:b547722454 "2"^^ . . _:b547722449 "2"^^ . _:b547722448 "2"^^ . _:b547722663 . _:b547722451 "2"^^ . _:b547722223 . _:b547722450 "2"^^ . _:b547722337 . _:b547722461 "2"^^ . _:b29675215 "Delineation of the mechanisms by which HBV evades host immunity to establish chronic infection and promote liver disease is hampered by the lack of robust animal models [11], [>>12<<], [13]." . _:b547722460 "2"^^ . . . _:b547722463 "2"^^ . . _:b547722462 "2"^^ . _:b547722457 "2"^^ . . _:b547722456 "2"^^ . _:b547722605 . _:b547722459 "2"^^ . _:b547722458 "2"^^ . _:b547722469 "2"^^ . _:b547722468 "2"^^ . . _:b547722471 "2"^^ . _:b547722470 "2"^^ . _:b547722457 . _:b547722465 "2"^^ . . . _:b547722464 "2"^^ . _:b29675286 _:b29675308 . _:b29675286 _:b29675309 . _:b547722467 "2"^^ . _:b29675286 _:b29675304 . _:b29675286 _:b29675305 . _:b29675286 _:b29675306 . _:b29675286 _:b29675307 . _:b547722466 "2"^^ . _:b29675286 _:b29675300 . _:b29675286 _:b29675301 . _:b29675286 _:b29675302 . _:b547722557 . _:b547722601 . _:b29675286 _:b29675303 . _:b547722477 "2"^^ . _:b29675286 _:b29675296 . _:b29675286 _:b29675297 . _:b29675286 _:b29675298 . . _:b29675286 _:b29675299 . _:b547722476 "2"^^ . _:b29675286 _:b29675292 . _:b29675286 _:b29675293 . _:b29675286 _:b29675294 . . _:b29675286 _:b29675295 . _:b547722479 "2"^^ . _:b29675286 _:b29675288 . _:b29675286 _:b29675289 . _:b29675286 _:b29675290 . _:b29675286 _:b29675291 . _:b547722478 "2"^^ . . _:b29675286 _:b29675287 . _:b547722246 . _:b547722473 "2"^^ . _:b547722472 "2"^^ . _:b547722137 . _:b547722475 "2"^^ . _:b29675233 . _:b547722474 "2"^^ . . _:b547722485 "2"^^ . _:b547722484 "2"^^ . . _:b547722487 "2"^^ . . . _:b547722486 "2"^^ . . _:b547722481 "2"^^ . _:b547722578 . _:b547722480 "2"^^ . . _:b547722507 . _:b547722483 "2"^^ . . _:b547722452 . _:b547722661 . _:b547722482 "2"^^ . _:b547722493 "2"^^ . _:b547722273 . _:b547722492 "2"^^ . _:b29675264 _:b29675268 . _:b547722473 . _:b29675294 . _:b29675264 _:b29675269 . _:b29675264 _:b29675270 . _:b29675264 _:b29675271 . _:b547722495 "2"^^ . _:b29675264 _:b29675265 . _:b29675264 _:b29675266 . . . _:b29675264 _:b29675267 . _:b547722494 "2"^^ . _:b29675264 _:b29675276 . _:b29675264 _:b29675277 . _:b29675264 _:b29675278 . _:b29675264 _:b29675279 . . _:b29675209 "Though HBV-induced liver disease is predominately a chronic disease, requiring decades of chronic infection and liver inflammation [5], [>>6<<], [7], HBV infection occasionally results in accelerated liver disease and liver failure during acute infection [8], [9]." . _:b29675264 _:b29675272 . _:b547722489 "2"^^ . _:b29675264 _:b29675273 . _:b29675264 _:b29675274 . _:b29675264 _:b29675275 . _:b547722488 "2"^^ . _:b29675264 _:b29675284 . _:b29675264 _:b29675285 . _:b547722568 . _:b547722491 "2"^^ . _:b29675264 _:b29675280 . _:b547722264 . _:b29675264 _:b29675281 . _:b29675264 _:b29675282 . _:b29675264 _:b29675283 . _:b547722490 "2"^^ . _:b547722501 "2"^^ . _:b29675279 "gene expression profile (CD68high, CD163high, AMAC1high, iNOSlow, TNF\u03B1low) is associated with upregulation of tissue fibrosis (COL1A1high, TIMP1high) and damage markers (HMGB1high [42], [43], [44], S100A9high [45], [46]) (Figure S9) [>>8<<], [9]." . . _:b547722500 "2"^^ . . _:b547722503 "2"^^ . _:b547722502 "2"^^ . _:b547722384 . _:b547722497 "2"^^ . . _:b29675231 . _:b29675275 "profile analysis showed M2-like macrophage gene expression profile (CD68high, CD163high, AMAC1high, iNOSlow, TNF\u03B1low) is associated with upregulation of tissue fibrosis (COL1A1high, TIMP1high) and damage markers (HMGB1high [42], [>>43<<], [44], S100A9high [45], [46]) (Figure S9) [8], [9]." . _:b547722496 "2"^^ . . _:b547722499 "2"^^ . _:b29675300 "In chronically infected patients, immune and inflammatory responses against HBV are implicated as the major mediators of liver diseases [54], [>>55<<]. Chronic HBV infection in the liver of A2/NSG/Fas-hu mice was associated with significant human leukocyte infiltration, leading to human hepatic stellate cell activation and human liver fibrosis." . . _:b547722498 "2"^^ . . _:b547722235 . _:b547722509 "2"^^ . . _:b547722508 "2"^^ . . . _:b547722511 "2"^^ . . . _:b547722510 "2"^^ . _:b547722322 . _:b547722505 "2"^^ . _:b547722131 . _:b547722504 "2"^^ . _:b547722507 "2"^^ . . _:b547722506 "2"^^ . . _:b547722517 "2"^^ . _:b547722516 "2"^^ . . _:b547722519 "2"^^ . _:b29675256 "Immunoreactivity was determined by incubation with DAB substrate (Pierce) or Vulcan red (Dako), and counterstained with hematoxilin [>>56<<], [58]. Liver histological activity was determined using the knodell score, which examines liver necrosis, degeneration, inflammation and fibrosis [62], [63]." . . _:b547722518 "2"^^ . _:b547722383 . . _:b547722513 "2"^^ . _:b547722530 . _:b547722512 "2"^^ . _:b29675232 . _:b547722515 "2"^^ . _:b547722402 . _:b547722514 "2"^^ . _:b29675244 "To promote human liver cell repopulation, A2/NSG-hu HSC/Hep mice were treated with a murine specific anti-Fas agonistic antibody (Jo2) [32], [33], [34], [>>35<<], [36]. The A2/NSG-hu HSC/Hep mouse model enabled human liver and immune system development and supported long-term HBV infection, anti-HBV human immune response and HBV-induced liver diseases including hepatitis and fibrosis." . _:b547722525 "2"^^ . _:b547722524 "2"^^ . _:b547722527 "2"^^ . _:b547722526 "2"^^ . _:b29675302 "and associated tissue fibrosis and damage; with M1 polarized macrophages promoting anti-virus Th1 immune response and pathogen clearance, while M2 polarized macrophages impair Th1 immune response and promoting tissue remodeling [23], [>>24<<], [25], [26], [27]." . _:b547722521 "2"^^ . _:b547722520 "2"^^ . _:b547722472 . _:b547722523 "2"^^ . . _:b547722380 . _:b547722522 "2"^^ . _:b547722217 . _:b547722305 . _:b547722533 "2"^^ . _:b547722339 . _:b29675231 "clearance, chronic inflammation and associated liver pathology; with M1 polarized macrophages promoting pathogen clearance, and M2-like polarized macrophages impairing host immunity and promoting tissue fibrosis/remodeling [23], [>>24<<], [25], [26], [27]." . _:b547722652 . _:b547722532 "2"^^ . . _:b547722535 "2"^^ . _:b547722534 "2"^^ . _:b547722529 "2"^^ . _:b547722141 . . _:b547722528 "2"^^ . _:b29675298 . _:b29675304 . _:b547722531 "2"^^ . _:b547722530 "2"^^ . _:b29675225 "To overcome the limitations associated with current chimeric human-murine liver mouse models, we have recently developed a humanized mouse model with both human immune system and liver cells (AFC8-hu HSC/Hep mice) [18], [>>19<<]. AFC8-hu HSC/Hep mice can support HCV infection in the liver and generate human T-cell response to HCV. Additionally, HCV infection induces liver inflammation and fibrosis, correlated with activation of human hepatic stellate cells and" . . _:b547722541 "2"^^ . . . . _:b547722540 "2"^^ . _:b547722543 "2"^^ . _:b547722542 "2"^^ . . _:b29675225 . _:b547722537 "2"^^ . . _:b547722536 "2"^^ . _:b29675268 . . _:b547722539 "2"^^ . _:b547722241 . _:b547722112 . . _:b547722538 "2"^^ . . _:b547722549 "2"^^ . _:b547722548 "2"^^ . _:b547722336 . _:b547722551 "2"^^ . _:b547722550 "2"^^ . _:b547722545 "2"^^ . . _:b547722544 "2"^^ . . _:b547722547 "2"^^ . _:b547722546 "2"^^ . _:b547722557 "2"^^ . _:b547722144 . . _:b547722191 . _:b547722556 "2"^^ . _:b547722559 "2"^^ . _:b547722594 . _:b547722558 "2"^^ . . . _:b547722553 "2"^^ . . _:b547722552 "2"^^ . . _:b547722555 "2"^^ . . _:b547722554 "2"^^ . . _:b547722565 "2"^^ . _:b547722319 . _:b547722564 "2"^^ . _:b547722363 . _:b547722567 "2"^^ . . _:b547722566 "2"^^ . _:b547722434 . . _:b547722561 "2"^^ . . _:b547722560 "2"^^ . _:b547722563 "2"^^ . _:b547722651 . _:b547722514 . _:b547722562 "2"^^ . . _:b547722136 . _:b547722573 "2"^^ . _:b547722437 . _:b547722263 . . _:b29675252 "Animals were injected 3\u20135 times via ip with Jo2 antibody/PBS at 0.1\u20130.15 mg/kg body weight (BD Pharmingen) every 4\u20135 days at approximately 3\u20134 weeks post transplant of human cells [>>59<<]. At 12\u201316 weeks post-transplant with HSC+Hep cells, Transplanted mice were bled to determine human leukocyte (hCD45+) reconstitution by FACS and human albumin concentration in the blood by ELISA (Bethyl laboratories). All experiments" . _:b547722572 "2"^^ . _:b547722575 "2"^^ . _:b547722272 . _:b547722574 "2"^^ . . _:b547722569 "2"^^ . _:b547722568 "2"^^ . _:b547722571 "2"^^ . . _:b547722570 "2"^^ . _:b547722649 . _:b547722581 "2"^^ . . _:b547722580 "2"^^ . _:b547722583 "2"^^ . . _:b547722582 "2"^^ . _:b547722577 "2"^^ . . _:b547722576 "2"^^ . . _:b547722407 . _:b547722579 . _:b547722579 "2"^^ . . _:b547722578 "2"^^ . _:b547722589 "2"^^ . . _:b547722588 "2"^^ . . _:b547722591 "2"^^ . . . _:b547722590 "2"^^ . _:b547722591 . _:b547722155 . _:b547722585 "2"^^ . _:b547722504 . . _:b547722584 "2"^^ . _:b547722152 . . _:b547722587 "2"^^ . _:b29675213 "However, HBV is endemic in many developing countries with over 350 million people worldwide chronically infected [>>10<<]. Delineation of the mechanisms by which HBV evades host immunity to establish chronic infection and promote liver disease is hampered by the lack of robust animal models [11], [12], [13]." . _:b547722312 . _:b547722586 "2"^^ . . _:b547722392 . _:b547722597 "2"^^ . _:b547722373 . . _:b547722596 "2"^^ . _:b547722599 "2"^^ . . _:b547722598 "2"^^ . . _:b547722593 "2"^^ . . _:b547722592 "2"^^ . . _:b547722427 . _:b547722595 "2"^^ . . _:b547722594 "2"^^ . _:b29675280 "expression profile (CD68high, CD163high, AMAC1high, iNOSlow, TNF\u03B1low) is associated with upregulation of tissue fibrosis (COL1A1high, TIMP1high) and damage markers (HMGB1high [42], [43], [44], S100A9high [45], [46]) (Figure S9) [8], [>>9<<]." . . _:b547722605 "2"^^ . . _:b547722604 "2"^^ . _:b547722607 "2"^^ . _:b547722606 "2"^^ . _:b547722601 "2"^^ . _:b29675281 "Results from several cell culture studies have suggested that HBV can modulate monocyte activation resulting in induction of M2 associated cytokines and inhibition of M1 associated cytokines [>>20<<], [22], [47], [48], [49]." . _:b547722600 "2"^^ . _:b547722603 "2"^^ . _:b547722602 "2"^^ . . _:b547722613 "2"^^ . . _:b547722147 . _:b547722612 "2"^^ . _:b547722615 "2"^^ . _:b547722547 . . _:b547722614 "2"^^ . _:b547722498 . _:b547722609 "2"^^ . _:b547722250 . . _:b547722106 . _:b547722608 "2"^^ . . . _:b547722611 "2"^^ . _:b547722610 "2"^^ . _:b547722259 . _:b547722621 "2"^^ . . _:b547722620 "2"^^ . _:b547722417 . _:b547722623 "2"^^ . _:b547722622 "2"^^ . . _:b547722369 . _:b547722603 . _:b547722174 . _:b547722617 "2"^^ . _:b29675219 "Additionally, the fumarylacetoacetate hydrolase (Fah)-Rag2-\u03B3C-null mice also allow human hepatocytes engraftment and HCV infection [>>15<<], [16], [17]." . . _:b547722616 "2"^^ . _:b547722619 "2"^^ . . _:b29675233 "chronic inflammation and associated liver pathology; with M1 polarized macrophages promoting pathogen clearance, and M2-like polarized macrophages impairing host immunity and promoting tissue fibrosis/remodeling [23], [24], [25], [>>26<<], [27]." . _:b547722169 . _:b547722618 "2"^^ . . _:b547722460 . _:b547722629 "2"^^ . . _:b547722628 "2"^^ . _:b547722631 "2"^^ . _:b547722630 "2"^^ . _:b29675267 "Chronic HBV infection in patients is associated with chronic hepatitis and liver fibrosis, characterized by leukocyte infiltration and collagen deposition in portal/periportal regions of the liver [>>38<<]. To examine leukocyte infiltration and fibrosis in HBV infected liver of humanized mice, liver sections were examined at time of sacrifice." . _:b547722625 "2"^^ . _:b547722167 . _:b547722624 "2"^^ . . _:b547722627 "2"^^ . . . _:b547722626 "2"^^ . _:b547722637 "2"^^ . _:b547722636 "2"^^ . . _:b29675209 . _:b547722639 "2"^^ . . . . _:b547722638 "2"^^ . _:b547722495 . _:b547722646 . _:b547722633 "2"^^ . _:b547722632 "2"^^ . _:b547722501 . _:b547722635 "2"^^ . _:b547722500 . _:b547722634 "2"^^ . _:b547722503 . _:b29675299 . . _:b547722645 "2"^^ . _:b547722502 . _:b29675229 . _:b547722644 "2"^^ . _:b547722497 . _:b547722647 "2"^^ . _:b547722496 . _:b547722597 . _:b547722646 "2"^^ . _:b547722499 . . _:b547722641 "2"^^ . _:b547722498 . _:b547722279 . _:b547722640 "2"^^ . _:b547722509 . . _:b547722643 "2"^^ . _:b547722508 . _:b547722642 "2"^^ . _:b547722583 . _:b547722511 . _:b29675210 . _:b547722653 "2"^^ . _:b547722510 . _:b547722652 "2"^^ . _:b547722505 . . _:b547722655 "2"^^ . _:b547722504 . _:b547722665 . _:b547722654 "2"^^ . _:b547722507 . _:b29675306 . . _:b547722649 "2"^^ . _:b547722506 . . . _:b547722648 "2"^^ . _:b547722517 . _:b547722584 . _:b547722651 "2"^^ . _:b547722516 . . _:b547722650 "2"^^ . _:b547722519 . _:b29675258 . _:b547722661 "2"^^ . _:b547722518 . _:b547722662 . _:b547722566 . _:b547722655 . _:b547722323 . _:b547722660 "2"^^ . _:b547722513 . _:b547722256 . _:b29675298 "However, only suboptimal B cell response has been reported thus far in humanized mouse models [>>52<<]. In concordance with those studies, we detected predominantly human IgM antibodies with anti-HBV activity in HBV infected animals. Although human lymphoid T cells exhibited robust anti-HBV immune responses, HBV infection resulted in" . _:b547722350 . _:b547722663 "2"^^ . _:b547722512 . _:b547722519 . _:b547722662 "2"^^ . . _:b29675241 "To promote human liver cell repopulation, A2/NSG-hu HSC/Hep mice were treated with a murine specific anti-Fas agonistic antibody (Jo2) [>>32<<], [33], [34], [35], [36]." . _:b547722515 . . _:b547722520 . _:b547722657 "2"^^ . _:b547722514 . _:b547722656 "2"^^ . _:b547722525 . _:b547722659 "2"^^ . _:b547722524 . _:b547722395 . _:b547722107 . _:b547722658 "2"^^ . _:b547722527 . . _:b547722669 "2"^^ . _:b547722526 . . _:b29675234 "inflammation and associated liver pathology; with M1 polarized macrophages promoting pathogen clearance, and M2-like polarized macrophages impairing host immunity and promoting tissue fibrosis/remodeling [23], [24], [25], [26], [>>27<<]." . _:b547722419 . _:b547722425 . _:b547722668 "2"^^ . _:b547722521 . _:b29675283 . _:b547722499 . _:b547722671 "2"^^ . _:b547722520 . _:b547722400 . _:b547722311 . _:b547722670 "2"^^ . _:b547722523 . _:b547722665 "2"^^ . _:b547722522 . _:b547722522 . _:b29675303 "tissue fibrosis and damage; with M1 polarized macrophages promoting anti-virus Th1 immune response and pathogen clearance, while M2 polarized macrophages impair Th1 immune response and promoting tissue remodeling [23], [24], [>>25<<], [26], [27]." . _:b547722664 "2"^^ . _:b547722533 . . _:b547722115 . _:b547722626 . _:b547722667 "2"^^ . _:b547722532 . . _:b29675276 "profile analysis showed M2-like macrophage gene expression profile (CD68high, CD163high, AMAC1high, iNOSlow, TNF\u03B1low) is associated with upregulation of tissue fibrosis (COL1A1high, TIMP1high) and damage markers (HMGB1high [42], [43], [>>44<<], S100A9high [45], [46]) (Figure S9) [8], [9]." . _:b547722676 . _:b547722666 "2"^^ . _:b547722535 . _:b547722239 . _:b547722677 "2"^^ . _:b547722534 . _:b29675269 . _:b547722676 "2"^^ . _:b547722529 . _:b547722528 . _:b547722262 . _:b547722531 . . _:b29675248 "Liver gene expression profile analyses in patients were obtained from a dataset in Gene Expression Omnibus (GEO)/NCBI database; the reports followed NIH research ethics guidelines [8], [>>9<<]." . . _:b547722673 "2"^^ . _:b547722530 . _:b29675253 "HBV serum genome was detected at termination using real-time PCR [>>60<<]. Serum cytokine levels were measure using a multiplex human cytokine array and following manufacturer's recommended procedures (Luminex, Millipore)." . . _:b547722672 "2"^^ . _:b547722541 . _:b547722675 "2"^^ . _:b547722540 . . . . . _:b547722674 "2"^^ . _:b547722543 . . _:b547722494 . _:b547722542 . _:b547722537 . . _:b29675245 "To promote human liver cell repopulation, A2/NSG-hu HSC/Hep mice were treated with a murine specific anti-Fas agonistic antibody (Jo2) [32], [33], [34], [35], [>>36<<]. The A2/NSG-hu HSC/Hep mouse model enabled human liver and immune system development and supported long-term HBV infection, anti-HBV human immune response and HBV-induced liver diseases including hepatitis and fibrosis. Interestingly, we" . _:b547722536 . . _:b547722539 . _:b547722468 . _:b547722538 . . _:b547722549 . _:b547722548 . _:b547722163 . _:b547722551 . _:b547722550 . _:b547722286 . _:b547722575 . _:b547722545 . . . _:b547722544 . . _:b547722547 . _:b547722153 . _:b547722546 . . _:b547722557 . . _:b547722556 . . _:b547722612 . _:b547722559 . . _:b547722558 .