Rejection of Experimental Hodgkins Lymphoma by T-Cells Engineered with a CD19 Chimeric Antigen Receptor

T cells engineered to express chimeric antigen receptors (CARs) combining an external antibody binding domain with the CD3ζ T cell receptor (TCR) signaling domain for triggering cell activation are being used for immunotherapeutic targeting of tumor cells in a non-HLA restricted manner. In this study we transduced T cells with a CD19-CAR construct containing a truncated CD34 gene (tCD34) marker and used these to target the B cell antigen CD19 on the surface of a Hodgkin’s lymphoma (HL) cell line (L591) both in vitro and in vivo. Levels of tCD34 expression in transduced peripheral blood mononuclear cells (PBMCs) ranged from 6% - 20% and this was increased to 82% after selection for transduced tCD34+ cells. In vitro cytotoxicity testing on a CD19+ HL cell line (L591) showed specific cell lysis initiated by the CD19-CAR transduced PBMCs. Importantly, CD19-CAR T cells prevented the growth of L591 HL tumor cells when co-injected subcutaneously (sc) in 6/6 severe combined immunodeficient (SCID) mice. There was no evidence of anti-tumor activity when CD19-CAR T cells were infused intravenously (iv) at the same time as L591 HL tumor cells were injected sc. However, 3/6 SCID mice showed tumor rejection within 83 days after iv infusion of CD19-CAR T cells 3 - 9 days after establishment of L591 HL tumors, while all control animals succumbed to tumors within 60 days. Interestingly, immuno-histochemical analysis of L591 HL tumors demonstrated that CD19-CAR T cells were detected not earlier than 11 days after infusion within the tumor mass. These results suggest that CD19 is a potentially attractive target for the immunotherapy of HL.

成红细胞样Ter细胞在胶原诱导性关节炎发病中的作用

目的:通过检测Ter细胞在不同发病阶段胶原诱导性关节炎(collagen-induced arthritis,CIA)小鼠脾脏中的数量变化及高峰期CIA小鼠脾脏Ter细胞与关节评分和T、B细胞亚群比例的相关性,探讨Ter细胞在CIA发生和发展中的作用,从而进一步深入理解类风湿关节炎的发病机制。方法: 6~8周的DBA/1小鼠进行CIA模型的诱导,二次免疫后开始对CIA小鼠进行关节评分。根据发病时间和关节评分将CIA分为发病早期、高峰期、晚期三个阶段。发病高峰期小鼠根据最终关节评分再分为高评分组(>8分)和低评分组(≤8分),流式细胞术检测na ve小鼠和各个阶段CIA小鼠脾脏中Ter细胞比例及发病高峰期CIA小鼠脾脏T、B 细胞亚群比例,并进行关联分析。结果:发病高峰期CIA小鼠脾脏Ter细胞比例较na ve小鼠明显升高(8.522%±2.645% vs . 1.937%±0.725%, P < 0.01),高评分组小鼠脾脏Ter细胞比例明显低于低评分组(6.217%±0.841% vs . 10.827%±0.917%, P < 0.01 )。高评分组小鼠脾脏Th1细胞比例明显高于低评分组(1.337%±0.110% vs . 0.727%±0.223%, P < 0.05 ),高评分组小鼠脾脏Th17细胞比例高于低评分组(0.750%±0.171% vs . 0.477%±0.051%, P =0.099),高评分组小鼠脾脏生发中心B(germinal center B,GC-B)细胞比例明显高于低评分组(1.243%±0.057% vs . 1.097%±0.015%, P < 0.05 )。相关性分析结果显示,发病高峰期CIA小鼠脾脏Ter细胞比例与CD4 + T、Th1、Th17、GC-B细胞比例均呈强负相关,与B10细胞比例呈强正相关,相关性均有统计学意义,提示这群细胞在CIA中可能具有保护作用。动态变化研究显示,随着疾病进展,发病晚期CIA小鼠脾脏Ter细胞比例较高峰期明显降低(0.917%± 0.588% vs . 8.522%±2.645%, P <0.001),进一步提示这群细胞在关节炎中的保护作用。结论: Ter细胞在发病高峰期CIA小鼠脾脏中明显增加,与小鼠关节评分及致病性免疫细胞比例呈负相关,与保护性免疫细胞比例呈正相关,发病晚期CIA小鼠Ter细胞比例明显降低,提示Ter细胞可能作为一种保护性细胞参与类风湿关节炎的发生和发展,但其具体作用及机制仍需后续进一步的体内及体外实验验证。

Cytokines as critical co-stimulatory molecules in modulating the immune response of natural killer cells

Cytokines are involved in directing the activation of natural killer （NK） cells. NK cells are involved in the recognition of cells that have been altered; thus they do not recognize specific insults to the host, but when activated, are capable of destroying infected cells directly, as well as promoting the recruitment and response of the other components of the immune system by the release of cytokines and chemokines. It is these properties that have made NK cells a critical part of innate immunity and adaptive immunity, and they play a principal role linking innate and adaptive immunity by the recruitment of an adaptive immune response to an innate immune reaction.

Clusterin mRNA expression in apoptotic and activated rat thymocytes

Clusterin is a 75-80 kDa heterodimeric glycoprotein, that is produced in most tissues but which exactbiological role is still not clear. Particularly, its role in protection or promotion of apoptosis is heavilydisputed, since data supporting both views have been reported in several independent studies. To clarify thisissue, and also to determine whether clusterin expression itself might be affected by apoptosis, in the presentstudy, rat thymocytes were treated with dexamethasone, -a synthetic glucocorticoid that elicits apoptosis inthymocytes-, and clusterin mRNA expression was analyzed by semi-quantitative RT-PCR before and afterinduction of apoptosis. Interestingly, neither the treatment with dexamethasone in vitro nor triggering ofapoptosis in vivo up- regulated clusterin expression, opposing the view that clusterin is involved in apoptoticprocesses. On the other hand, a new clusterin mRNA isoform was detected and isolated, whose expressionwas restricted to freshly isolated thymocytes. This novel isoform lacks the post-translational proteolyticcleavage site and is therefore predicted to encode a monomeric protein. The biological function undernormal circumstances, however, will need further investigations for clarification. While apoptosis could notmodulate clusterin expression, activation of thymocytes with concanavalin A and interleukin-2 resulted inup-regulation of clusterin mRNA level, indicating that clusterin expression is rather under the control ofcell activation-mediated rather than apoptosis- induced signals.

Tumor microenvironment in primary liver tumors: A challenging role of natural killer cells

In the last years,several studies have been focused on elucidate the role of tumor microenvironment(TME)in cancer development and progression.Within TME,cells from adaptive and innate immune system are one of the main abundant components.The dynamic interactions between immune and cancer cells lead to the activation of complex molecular mechanisms that sustain tumor growth.This important cross-talk has been elucidate for several kind of tumors and occurs also in patients with liver cancer,such as hepatocellular carcinoma(HCC)and intrahepatic cholangiocarcinoma(iCCA).Liver is well-known to be an important immunological organ with unique microenvironment.Here,in normal conditions,the rich immune-infiltrating cells cooperate with non-parenchymal cells,such as liver sinusoidal endothelial cells and Kupffer cells,favoring self-tolerance against gut antigens.The presence of underling liver immunosuppressive microenvironment highlights the importance to dissect the interaction between HCC and iCCA cells with immune infiltrating cells,in order to understand how this cross-talk promotes tumor growth.Deeper attention is,in fact,focused on immune-based therapy for these tumors,as promising approach to counteract the intrinsic anti-tumor activity of this microenvironment.In this review,we will examine the key pathways underlying TME cell-cell communications,with deeper focus on the role of natural killer cells in primary liver tumors,such as HCC and iCCA,as new opportunities for immune-based therapeutic strategies.

Analysis of peripheral blood dendritic cells as a non-invasivetool in the follow-up of patients with chronic hepatitis C

Hepatitis C virus(HCV) has a high propensity to establish chronic infections. Failure of HCV-infected individuals to activate effective antiviral immune responses is at least in part related to HCV-induced impairment of dendritic cells(DCs) that play a central role in activating T cell responses. Although the impact of HCV on DC phenotype and function is likely to be more prominent in the liver, major HCV-induced alterations are detectable in peripheral blood DCs(pb DCs) that represent the most accessible source of DCs. These alterations include numerical reduction, impaired production of inflammatory cytokines and increased production of immunosuppressive IL10. These changes in DCs are relevant to our understanding the immune mechanisms underlying the propensity of HCV to establish persistent infection. Importantly, the noninvasive accessibility of pb DCs renders the analysis of these cells a convenient procedure that can be serially repeated in patient follow-up. Accordingly, the study of pb DCs in HCV-infected patients during conventional treatment with pegylated interferon and ribavirin indicated that restoration of normal plasmacytoid DC count may represent an additional mechanism contributing to the efficacy of the dual therapy. It also identified the pre-treatment levels of plasmacytoid DCs and IL10 as putative predictors of response to therapy. Treatment of chronic HCV infection is changing, as new generation direct-acting antiviral agents will soon be available for use in interferon-free therapeutic strategies. The phenotypic and functional analysis of pb DCs in this novel therapeutic setting will provide a valuable tool for investigating mechanisms underlying treatment efficacy and for identifying predictors of treatment response.

Regulation of neuroinflammatory properties of glial cells by T cell effector molecules

Multiple sclerosis （MS） is a chronic inflammatory and neurodegenerative disorder that is thought to be mediated by autoreactive T lymphocytes that find their way into the central nervous system （CNS）. The pathological mechanism of MS is still being elucidated but it involves complex interactions between infiltrating immune cells and resi- dent glial cells within the CNS that culminate into strong neuroinflammation and axonal damage.

抗白细胞介素-12单克隆抗体阻断慢性实验性结肠炎发生

目的 探讨抗白介素 12 (IL 12 )单克隆抗体治疗实验性慢性结肠炎的机制。方法 用严重联合免疫缺陷 (SCID)小鼠 ,植入同基因源性CD4 5RBhighCD+4 T细胞 ,诱导慢性结肠炎 ,并进行抗IL 12单克隆抗体治疗和观察IL 12在慢性结肠炎中的病理生理作用。结果 在T细胞植入 4周后结肠组织内IL 12P4 0 mRNA含量明显升高 ,抗IL 12单克隆抗体有效地阻断了慢性结肠炎的发生 ,降低了结肠粘膜内CD4 +T细胞和巨噬细胞浸润 ,并抑制肠粘膜内固有层CD4 +T细胞分泌IL 2和IFN γ。结论 IL 12参与了慢性实验性结肠炎的病理生理过程 ,阻断IL

The talented interferon-gamma

IFN-γ is an extraordinarily pleotropic cytokine. It can not only heighten both the innate and adaptive immune response against pathogens and tumors, but also has the ability to maintain immune homeostasis. Since the effects of IFN-γ are cell and tissue specific, it is important to consider the recent advances in IFN-γ signaling in the context of different diseases. To this end, we review the involvement of IFN-γ in the pathogenesis of several inflammatory diseases, its therapeutic potential as an anti-tumor agent and its effects upon stem cells.

Comparable outcomes but higher risks of prolonged viral RNA shedding duration and secondary infection in cancer survivors with COVID-19: A multi-center, matched retrospective cohort study

Results Sixty-one cancer survivors and 183 matched non-cancer patients were screened from 2,828 COVID-19 infected patients admitted to 4 hospitals in Wuhan,China.The median ages of the cancer survivor cohort and non-cancer patient cohort were 64.0(55.0–73.0)and 64.0(54.0–73.5),respectively(P=0.909).Cancer survivors reported a higher incidence of symptom onset than non-cancer patients.Fever(80.3%vs.65.0%;P=0.026)was the most prevalent symptom,followed by cough(65.6%vs.37.7%;P<0.001),myalgia,and fatigue(45.9%vs.13.6%;P<0.001).The risks of the development of severe events(adjusted hazard ratio[AHR]=1.25;95%confidence interval[CI]:0.76–2.06;P=0.378)and mortality(relative risk[RR]=0.90,95%CI:0.79–1.04;P=0.416)in the cancer survivor cohort were comparable to those of the matched non-cancer patient cohort.However,the cancer survivor cohort showed a higher incidence of secondary infection(52.5%vs.30.1%;RR=1.47,95%CI:1.11–1.95;P=0.002)and a prolonged viral RNA shedding duration(32 days[IQR 26.0–46.0]vs.24.0 days[IQR 18.0–33.0];AHR=0.54;95%CI:0.38–0.80;P<0.05).Conclusion Compared to non-cancer patients,cancer survivors with COVID-19 exhibited a higher incidence of secondary infection,a prolonged period of viral shedding,but comparable risks of the development of severe events and mortality.It is helpful for clinicians to take tailored measures to treat cancer survivors with COVID-19.

Favorable effect of modest alcohol consumption to fatty liver disease

We previously reported that modest alcohol consumption was significantly inversely associated with fatty liver disease.Feng et al pointed out a discrepancy of statistical significance between our current larger scale cohort and a previous cohort.However,the prevalence of non-alcoholic fatty liver disease was higher in non or minimal drinkers than those in light drinkers in both cohorts.They also argue that some potential co-factors such as soft drink consumption and genetic variations should be discussed.

Projection of an immunological self shadow to developing T cells via macroautophagy

The adaptive immune system, which establishes specific memory for pathogens and foreign antigens that we encounter, is orchestrated by T cells. In contrast to antibody producing B cells, T cells recognize their antigen not in its native form, but as small proteolytic fragments presented by major histocom-patibility （MHC） molecules on the cell surface.

Comprehensive Analysis of Key mRNAs and lncRNAs in Osteosarcoma Response to Preoperative Chemotherapy with Prognostic Values

Objective:Osteosarcoma is one of the most common types of bone sarcoma with a poor prognosis.However,identifying the predictive factors that contribute to the response to neoadjuvant chemotherapy remains a significant challenge.Methods:A public data series(GSE87437)was downloaded to identify differentially expressed genes(DEGs)and differentially expressed lncRNAs(DElncRNAs)between osteosarcoma patients that do and do not respond to preoperative chemotherapy.Subsequently,functional analysis of the transcriptome expression profile,regulatory networks of DEGs and DElncRNAs,competing endogenous RNAs(ceRNA)and protein-protein interaction networks were performed.Furthermore,the function,pathway,and survival analysis of hub genes was performed and drug and disease relationship prediction of DElncRNA was carried out.Results:A total of 626 DEGs,26 DElncRNAs,and 18 hub genes were identified.However,only one gene and two lncRNAs were found to be suitable as candidate gene and lncRNAs respectively.Conclusion:The DEGs,hub genes,candidate gene,and candidate lncRNAs screened out in this context were considered as potential biomarkers for the response to neoadjuvant chemotherapy of osteosarcoma.

Multiple sclerosis:why we should focus on both sides of the(auto)antibody

Various clinical and experimental findings suggest a pathogenic role of antibodies in multiple sclerosis(MS).Yet,whether antibodies contribute to the pathogenesis or progression of MS is still a subject of intense debate.This controversy particularly results from unclarity regarding the target antigens of the antibodies that are found in the central nervous system(CNS)of MS patients.

<i>In vitro</i>activity and function of B7-H4-Ig fusion protein

B7-H4 has been shown to inhibit T cell proliferation, cytokine production and cell cycle in vitro. B7-H4 deficient mice develop exacerbated disease in the mouse models of Rheumatoid Arthritis (RA), Type 1 Diabetes (T1D) and Experimental Autoimmune Encephalomyelitis (EAE). On the other hand, B7-H4-Ig fusion protein has been documented to assuage the symptoms in mouse models of RA, T1D, and multiple sclerosis in vivo. In the present study, B7-H4-Ig bound to the majority of human peripheral blood monocytes and NK cells, but not to either normal or activated T cells. B7-H4-Ig fusion protein was assayed for its effects in allogeneic mixed lymphocyte culture (MLC) systems. Soluble B7- H4-Ig had no significant effect in the MLC, but with a tendency to promote allogeneic response. Immobilized, but not soluble B7-H4-Ig inhibited plastic bound anti-CD3 mediated activation of T cells. This inhibition however was largely due to B7-H4-Ig mediated displacement of anti-CD3 antibody from the plastic plate. Finally, B7-H4-Ig had no effect on the cytotoxicity mediated by NK and LAK cells in PBMC. Our findings thus caution against the interpretation of suppressive effect observed solely in plate-bound anti-CD3 mediated T cell co-stimulation in vitro.

Intravenous immunoglobulin suppresses IL-10 production by activated B cells in vitro

A therapeutic preparation of polyclonal human IgG, i.e., intravenous immunoglobulin (IVIg), has been employed to treat several inflammatory and autoimmune disorders. B cells are supposed to be a target of IVIg, but the molecular mechanism is elusive because of the lack of a suitable experimental system. To gain an insight into the beneficial effect of IVIg on B cells, we first established an experimental setting in which IVIg modulates a murine B cell function in vitro, and then aimed at identifying the mechanistic features at the molecular level. Here we show that IVIg down-regulates IL-10 production by CpG-activated B cells in vitro. The responsible component of IVIg was identified as the F(ab’)2 portion, whose polyclonality is mandatory for the suppressive effect. IVIg, bound to the surface of activated B cells, was found to be co-localized with intracellular SHP-1 on confocal laser microscopy, suggesting that B cell-surface immunoreceptor tyrosine-based inhibitory motif-harboring receptors that recruit SHP-1 are target molecules for IVIg in our experimental setting. Overall, we postulate a scenario in which IVIg attenuates B cells by suppressing IL-10 production, a B cell growth factor, and thus down-regulates the production of pathogenic antibodies.

Methodical and pre-analytical characteristics of a multiplex cancer biomarker immunoassay

AIM: To test the methodical and pre-analytical performance of a new multiplex cancer biomarker panel using magnetic beads. METHODS: The MILLIPLEX? MAP Human Circulating Cancer Biomarker Magnetic Bead Panel 1 comprises the tumor markers carcinoembryonic antigen, alpha-fetoprotein, total prostate-specific antigen, cancer antigen 15-3, cancer antigen 19-9, cancer antigen 125, cytokeratine 19-fragment, β-human chorionic gonadotropin, human epididymis protein 4, osteopontin, prolactin, the cell death and angiogenesis markers soluble Fas, soluble Fas-ligand, tumor necrosis factor related apoptosisinducing ligand, vascular endothelial growth factor andthe immunological markers interleukin-6(IL-6), IL-8, tumor necrosis factor-α, transforming growth factor α, fibroblast growth factor-2, macrophage migration inhibitory factor, leptin, hepatocyte growth factor, and stem cell factor. We determined intra- and inter-assay imprecision as well as dilution linearity using quality controls and serum pools. Furthermore, the stability of the 24 biomarkers examined in this panel was ascertained by testing the influence of different storage temperatures and time span before centrifugation.RESULTS: For all markers measured in the synthetic internal quality controls, the intra-assay imprecision ranged between 2.26% and 9.41%, while for 20 of 24 measured markers in the physiological serum pools, it ranged between 1.68% and 12.87%. The inter-assay imprecision ranged between 1.48%-17.12% for 23 biomarkers in synthetic, and between 4.59%-23.88% for 18 biomarkers in physiological quality controls. Here, single markers with very low concentration levels had increased imprecision rates. Dilution linearity was acceptable(70%-130% recovery) for 20 biomarkers. Regarding pre-analytical influencing factors, most markers were stable if blood centrifugation was delayed or if serum was stored for up to 24 h at 4 ℃ and 25 ℃ after centrifugation. Comparable results were obtained in serum and plasma for most markers. However, great changes were observed for single markers.CONCLUSION: MILLIPLEX? MAP Human Circulating Cancer Biomarker Magnetic Bead Panel 1 assay is a stable and precise method for detection of most biomarkers included in the kit. However, single markers have to be interpreted with care.

Recurrent Spontaneous Abortion of Immune Origin and HLA Sensitization Immunotherapy

Introduction: Spontaneous abortion is defined as pregnancy loss before the twentieth week and Recurrent Spontaneous Abortion (RSA) is defined as at least three spontaneous and successive pregnancy losses in the same period. Among the different types of immunological causes, this study refers specifically to the alloimmune cause. Women with RSA of alloimmune cause share a greater number of Human Leukocyte Antigen (HLA) with their husbands, leading to the inhibition of the production of anti-paternal asymmetric blocking antibodies, which would protect embryonic cells. Objectives: To evaluate the effect of immunotherapy with paternal HLA-sensitizing mononuclear cells in cases of RSA through the positivity of the cross-match test and its efficacy in pregnancy success. Patients and Methods: Written consent was obtained, 12 couples with a history of RSA presenting negative cross-match were included in the study. Blood samples were collected from the couple for cross-matching and the separation of paternal mononuclear cells. Immunotherapy was performed with paternal mononuclear cells intradermally in the wives’ arms on day (D) 0, D15 and D30. After the third dose, a second evaluation of the cross- match tests was performed. Results and Discussion: The age of the wives ranged from 28 to 41 years, with a mean of 34.5 years. The twelve couples had a negative cross-match test (100%). Eleven couples (96.6%) tested positive in the cross- match test after immunotherapy. Of these, 10 (90.9%) had successful pregnancies. Immunotherapy with paternal mononuclear cells showed an excellent ability to sensitize the maternal immune system, with positive cross-match, resulting in a successful pregnancy.

Kinin B_2 receptor does not exert renoprotective effects on mice with glycerol-induced rhabdomyolysis

AIM： To investigate a potential protective role of the kinin B2 receptor in a glycerol-induced rhabdomyolysis mouse model. METHODS： We separated 28 C57Bl/6 male mice into 4 groups： untreated WT animals, untreated B2 knockout mice, glycerol-treated WT and glycerol-treated B2 knockout mice. Glycerol-treated animals received one intramuscular injections of glycerol solution （50% v/v, 7 mL/kg）. After 48 h, urine and blood samples were collected to measure creatinine and urea levels. Addi-tionally, kidney samples were extracted for histological evaluation, and the mRNA expression levels of kinin B1 and B2 receptors and infammatory mediators were measured by real-time polymerase chain reaction. RESULTS： Serum creatinine and urea levels showed differences between untreated wild-type and glycerol-treated wild-type mice （0.66 ± 0.04 vs 2.61 ± 0.53 mg/dL, P 〈 0.01; and 33.51 ± 2.08 vs 330.2 ± 77.7 mg/dL, P 〈 0.005）, and between untreated B2 knock-out mice and glycerol-treated knockout mice （0.56 ± 0.03 vs 2.23 ± 0.87 mg/dL, P 〈 0.05; and 42.49 ± 3.2 vs 327.2 ± 58.4 mg/dL, P 〈 0.01）, but there was no difference between the glycerol-treated wild-type and glycerol-treated knockout mice. Glycerol was able to in-duce a striking increase in kinin B2 receptor expression （〉 30 times, 31.34 ± 8.9） in kidney. Animals injected with glycerol had a higher degree of tubular injury than untreated animals. Wild-type and knockout mice treat-ed with glycerol intramuscularly present kidney injury, with impairment in renal function. However, B2 knock-out mice treated with glycerol did not show a different phenotype regarding kidney injury markers, when com-pared to the wild-type glycerol-treated group. CONCLUSION： We conclude that the kinin B2 receptordoes not have a protective role in renal injury.

Study on Antibodies to Liver Antigens in Chinese Patients with Different Liver Diseases

In order to observe several antibodies to liver antigens in Chinese patients with different liver diseases and to discuss the characteristics of the autoantibodies in autoimmune liver diseases, from 1412 patients, detected by indirect immunofluorescence (IIF) initially, 230 patients with abnormal ALT were chosen and divided into 5 groups: ① autoimmune diseases group, 42 cases: 18 with autoimmune hepatitis (AIH), 21 with primary biliary cirrhosis (PBC), 3 with primary sclerosing cholangitis(PSC). ② HAV group, 23 cases; ③ HBV group, 70 cases; ④ HCV group, 35 cases and ⑤ Non A-E group, 60 cases. First, ANA, AMA, SMA, liver-kidney microsomal antibody (LKM) and so on were tested by IIF.Then, LKM-1, liver cytosolic-1 (LC-1), soluble liver antigen/liver pancreas (SLA/LP) and subtype of AMA (M2) as well as ANA profile such as SS-A, SS-B and dsDNA were tested by Western blot and immunoblot strips assay, respectively. The results were that among 1412 cases, those diagnosed as AIH, PBC and PSC accounted for 12.7‰, 14.9‰ and 2.1‰, respectively, of the samples being tested. 2/230 with LKM-1 and 2/230 with SLA/LP were seen in individuals infected with AIH and HCV, respectively. All patients with PBC showed AMA and M2 antibodies. No specific ANA pattern was seen in AIH by IIF but anti-actin was only found in patients with AIH. In Non A-E group, four cases were positive of AMA and M2; three had high titer of SMA and other 4 had SS-A, SS-B or dsDNA antibodies, etc. It was concluded that the detection of anti-liver antigens, ANA profile and AMA subtypes were helpful for the diagnosis of autoimmune liver diseases and overlap syndromes. In patients with Non A-E hepatitis, the diagnosis of PBC or AIH should be taken into consideration.