Human leukocyte antigen DRB1 1302 protects against bile duct damage and portal lymphocyte infiltration in patients with chronic hepatitis C.

BACKGROUND/AIMS: To confirm the immune reaction of hosts in chronic hepatitis C, we examined the association of human leukocyte antigen (HLA) DR with the histopathological outcome including bile duct damage and steatosis, which are characteristic of hepatitis C virus (HCV) infection. METHODS: One hundred and fifty-five patients with chronic HCV infection were examined. The pathological appearance of liver biopsy specimens was evaluated by both Knodell's histological activity index and examination of bile duct damage and steatosis. HLA DRB1 was determined by the polymerase chain reaction sequence-specific oligonucleotide probe method. RESULTS: HLA DRB1 1302 was found with significantly higher frequency in patients without than with bile duct damage (34.8% vs. 4.7%, p=0.0001, p corrected by Bonferroni's inequality method=0.002). It was also found more frequently in patients without marked portal lymphocyte infiltration (28.6% vs. 7.7%, p=0.0015, p corrected by Bonferroni's method=0.03). HLA DRB1 1101 was found more frequently in patients without than with piecemeal necrosis (p=0.004). In contrast, the frequency of HLA DRB1 1502 tended to be higher in patients with than without piecemeal necrosis and marked portal lymphocyte infiltration (p=0.015 and p=0.03, respectively). HLA DRB1 1201 and 0802 were seen more frequently in bile duct damage-negative (p=0.02) and piecemeal necrosis-negative patients (p=0.03), respectively. Interestingly, serum HCV levels of HLA DRB1 1302-positive patients were significantly higher than those of 1302-negative patients (mean: 7.7 Meq/ml vs. 3.1 Meq/ml, p=0.0007). CONCLUSION: These findings suggest that some histopathological changes in chronically HCV-infected livers could be caused by the host's immune reaction regulated by HLA DR.

HLA-DR and HLA-DQ gene typing of infertile women possessing sperm-immobilizing antibody.

Thirty-eight infertile women, possessing sperm-immobilizing antibody (SIA), were examined for their HLA-DR and -DQ types using DNA obtained from peripheral blood cells. The typing of HLA-DR and DQ was performed by polymerase chain reaction sequence specific oligonucleotide probes (PCR-SSOP) and PCR-restriction fragment length polymorphism (RFLP), respectively. In comparison to the normal Japanese population, the SIA positive patient population had higher genes frequencies in HLA-DRB1*0901 (26.3 vs. 13.6%, P<0.005), DQB1*0602 (13.2 vs. 6.2%, P<0.05) and -DQB1*0303 (26.3 vs. 14.8%, P<0.01), but not in any HLA-DQA1 gene types by chi2 test. After Bonferroni correction, the high frequency of HLA-DRB1*0901 remained significant (P<0.05) and HLA-DQB1*303 was slightly significant (P<0.07) but no other genes had a gene frequency significantly higher than that of the normal Japanese population. HLA-DRB1*0901 and HLA-DQB1*0303 are very rare among Caucasians but characteristically high among Japanese. The high frequency of HLA-DRB1*0901 and DQB1*0303 genes in the Japanese population may account for higher frequency of sperm-immobilizing antibody in Japanese compared to other ethnic groups.

Dominant and shared T cell receptor beta chain variable regions of T cells inducing synovial hyperplasia in rheumatoid arthritis.

Previously, we demonstrated the presence of at least two distinct subpopulations of patients with rheumatoid arthritis (RA) employing a cell-transfer experiment using severe combined immunodeficient (SCID) mice. One group of patients, whose T cells derived from the rheumatoid joints, induced synovial hyperplasia (SH) in the SCID mice (the positive group). The other group did not display the induction of SH (the negative group). TCR/Vbeta gene usage analysis indicated that some dominant T cell subpopulations were oligoclonally expanding only in the rheumatoid joints, and not in the periphery of the patients of the positive group. Moreover, these T cell subpopulations were not seen in the joints of patients in the negative group or in non-RA patients. In addition, the preferential uses of certain TCR/Vbetas (Vbeta8, Vbeta12, Vbeta13, and Vbeta14) genes were demonstrated in these T cells. In this study, to investigate whether these T cells are driven by a certain antigen(s), the third complementarity determining regions (CDR3s) of TCR/Vbeta, especially Vbeta8 and Vbeta14 PCR products, were cloned and sequenced. As a result, a dominant CDR3 sequence, CASS-PRERAT-YEQ, was found in Vbeta14+ T cells from the rheumatoid joint of a patient (Patient 1) of the positive group with a Vbeta14 skew. The identical CDR3 sequence also predominated in Vbeta14+ T cells from the rheumatoid joint of another patient (Patient 7) of the positive group with a Vbeta14 skew. In addition, in the patients (Patients 4, 7, 8) of the positive group with a Vbeta8 skew, other dominant CDR3 sequences, CASS-ENS-YEQ and CASS-LTEP-DTQ, were found as in the case of Vbeta14. However, no identical CDR3 sequences were detected dominantly in the joints of the patients in the negative group or in non-RA patients. A Vbeta14+ T cell clone (TCL), named G3, with the identical CDR3 sequence, CASS-PRERAT-YEQ, was isolated successfully from Patient 1, and cell transfer of G3 with autologous irradiated peripheral mononuclear cells induced SH in the SCID mice. Taken together, these results suggest that T cells inducing SH, thought to be pathogenic for RA, might be driven by a certain shared antigen(s).

[Clinical evaluation of enzyme immuno assay systems for HCV antibody--synthetic peptide antigen and recombinant antigen].

To evaluate the frequency of the false positive results, we examined 168 patients for the presence of HCV using two HCV antibody assay systems, Synpep HCV-EIA (Synpep) and Abbott EIA II (Abbott). The results obtained by the two methods were significantly different in 22 patients. Cases in 17 of these patients, the results were positive with Abbott but negative with Synpep, and there were no clinical signs or delectable virus RNA. However, in 2 cases, the results were markedly positive with Abbott and weakly positive with Synpep. The presence of virus RNA and the increase of transaminase were observed in one case but both were noted in the other case. The serum of these two patients reacted with the C33C and C22-3 regions in RIBA II. We observed another 2 cases in which the elevation of the cut-off index with Synpep preceded that with Abbott at the early stage of acute hepatitis C. We also compared the cut-off index with the histology activity index (HAI) score determined by liver biopsy. The average cut-off index with Synpep was proportional to the HAI score in the range between 0 and 13. Based on the cut-off index/HAI score relationship, we suggest that patients with inactive chronic hepatitis show a Synpep cut-off index less than 11.

T cell-mediated inhibition of the transfer of autoimmune diabetes in NOD mice.

The nonobese diabetic (NOD) mouse has recently been introduced as a model for insulin-dependent diabetes mellitus. The role of regulatory T cells in the development of antipancreatic autoimmunity in this model remains unclear. To evaluate the presence of suppressive phenomena, we used disease transfer by spleen cells from diabetic NOD mice into preirradiated adult recipients as a model for accelerated disease. Suppressor phenomena were detected by testing the protection afforded by lymphoid cells from nondiabetic NOD mice against diabetes transfer in irradiated recipients. Transfer of diabetes was delayed by reconstituting recipients with spleen cells from nondiabetic NOD donors. The greatest protection against diabetes transfer was conferred by spleen cells from 8-wk-old nondiabetic female NOD mice. Depletion experiments showed that the protection was dependent on CD4+ cells. Protection was also detected within thymic cells from nondiabetic NOD mice and protection conferred by spleen cells was abrogated by thymectomy of nondiabetic female, but not male, NOD donors at 3 wk of age. These findings indicate that suppressive CD4+ T cells that are dependent on the presence of the thymus may delay the onset of diabetes in female diabetes-prone NOD mice.

Anti-suppressor effect of cyclophosphamide on the development of spontaneous diabetes in NOD mice.

In the NOD mouse, an autoimmune process beginning by 5 weeks of age with lymphocyte infiltration and destruction of insulin-secreting beta cells leads to overt diabetes which begins to appear by 11 weeks of age. Although there is a high incidence of insulitis by 10 weeks of age (greater than 80%) in both males and females, by 30 weeks of age diabetic symptoms have occurred in 53-80% of females and in 12-40% of males. Intraperitoneal injection of a high dose (200 mg/kg) of cyclophosphamide (CY) consistently induces the onset of diabetes in male and female NOD mice at an age when spontaneous diabetes rarely occurs. Spleen T cells from CY-induced diabetic mice are capable of transferring the disease into irradiated nondiabetic syngeneic recipients. This indicates that the diabetogenic effect of CY is not mediated by direct toxicity on pancreatic beta cells but is mediated by abrogation of a suppressor mechanism which may prevent activation of T cells responsible for the development of diabetes in the NOD mouse. Additionally, CY is only effective in NOD mice and not in F1 hybrids between NOD and other strains of mice. Thus, the potential beta cell aggressor mechanism is not present in these hybrids as it is in homozygous mice, which indicates that it is not under the control of dominant genes.

[Early induction of diabetes in NOD mice by streptozotocin]. / Induction précoce du diabète chez la souris NOD par la streptozotocine.

To clarify whether the non-obese diabetes prone (NOD) mouse has an unusual pancreatic sensitivity to damage, mice were administered streptozotocin in high dose (direct beta cell toxic) or multiple low-dose (autoimmune-insulitis generating) regimen. NOD mice were found to be less sensitive to the diabetogenic effects of high-dose streptozotocin than C57BL/6 mice, but were exquisitely responsive to the multiple low dose regimen when compared to C57BL/6 or C3H/HeJ mice. These results suggest that the basic defect in NOD mice resides in the immune system and that the NOD mouse may be a useful model to investigate the relationships between environmental factors and intrinsic genetic predisposition to diabetes.

Killing of tumor cells in vitro by macrophages from mice given injections of squalene-treated cell wall skeleton of Nocardia rubra.

Peritoneal exudate cells (PEC) harvested from mice after i.p. injection of squalene-treated cell wall skeleton of Nocardia rubra (N. rubra-CWS) demonstrated vigorous cytolytic activity in vitro toward tumor target cells. Fractionation of these PEC by adherence to plastic dishes showed that the cytolytic activity in PEC was associated with an adherent phagocytic cell. Induction of the cytolytic adherent PEC required an optimal dose of 50 micrograms N. rubra-CWS and i.p. injection. Cytolytic activity of N. rubra-CWS-induced adherent PEC was maximal after 5 days and fell steadily thereafter. Susceptible tumor targets included cells syngeneic, allogeneic, and xenogeneic to the effector cell source. In contrast, nonneoplastic xenogeneic cells were not affected by N. rubra-CWS-induced adherent PEC. The effector cells were not found in the spleen or peripheral lymph nodes. In addition, the cytolytic activity of N. rubra-CWS-induced adherent PEC was completely inhibited by treatment with antimacrophage serum and complement or carrageenan. Treatment with monoclonal anti-Thy 1.2 antibody and complement, however, did not affect the cytolytic activity of the adherent PEC. These features make it likely that N. rubra-CWS-induced cytolytic effector cells are macrophages.

Primary lung cancer producing alpha-fetoprotein: a case report.

A case of primary lung cancer that produced both alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) is presented. Alphafetoprotein in the supernatant of tissue extract from the primary lesion amounted to more than 40 microgram/ml, and its exact origin from tumor tissues was confirmed by the immunofluorescent study. We suppose that since the lung is one of the foregut derivatives it is not unlikely for lung cancer cells to produce AFP as well as CEA.