115 CYTOLOGICAL ANALYSIS OF HEPATIC GENE EXPRESSION AND IMMUNOLOGICAL RESPONSE OF MHC ANTIGENS IN MOUSE AMNIOTIC EPITHELIAL CELLS

Human amniotic epithelial cells (hAECs) have been reported to have unique properties. They express almost no class I and class II MHC antigens and lack response to interferon-γ (IFN-γ) which mediates the expression of those MHC molecules. Moreover, hAECs express some genes characteristic of hepatic cells. Therefore, hAECs seem to have multipotency and are expected to substitute for hepatic tissues in part. We aimed to develop the experimental model for investigating AECs in mice (mAECs). In this study, we examined the induction of MHC molecules by IFN-γ and the hepatic gene expression in mAECs. Murine amniotic membranes were collected from C57BL/6J females at 17.5 days of gestation. They were digested by 0.03% hyaluronidase followed by 0.2% collagenase treatment. Dissociated mAECs were cultured on dishes in DMEM supplemented with 10% FBS at 37°C under 5%CO2 in air. Embryonic fibroblasts (EFs) collected from C57BL/6J fetuses at 13.5 dpc were cultured in the same condition as mAECs. In Experiment I, the effect of IFN-γ on induction of MHC molecules in mAECs was examined. mAECs and EFs cultured in the presence or absence of IFN-γ at 1 × 103 U mL−1 for 72 h were recovered and incubated with FITC-conjugated antibodies against mouse H-2 MHC class I or I-A/I-E MHC class II antigens. The cells were analyzed by flow cytometry. In Experiment II, the expression of the genes in mAECs was examined by RT-PCR. mRNA was purified from adult liver, EFs, fresh mAECs, and mAECs cultured for 5 days. As the genes characteristic for hepatic cells, HNF-3α, HNF-3β, HNF-3γ, HNF-4, transthyretin (TTR), albumin, α-fetoprotein (AFP), glucose-6-phosphatase (G6P), and asialoglycoprotein receptor-1 (Asgr1) were examined. In Experiment I, cell-surface expression of class I and class II MHC antigens in response to IFN-γ was observed weakly in mAECs as compared to EFs, suggesting different property in hAECs which lack the expression of those antigens. In Experiment II, RT-PCR analysis showed that all of the genes except G6P were expressed in fresh mAECs. However, the expression of transcription factors such as HNF-3α, HNF-3β, HNF-4, and TTR, serum proteins such as albumin and AFP, and Asgr1 decreased after in vitro culture, contrary to the case of hAECs in which, for example, albumin appeared after cultivation. In conclusion, it was evident that mAECs have quite different properties, both in the inductivity of MHC molecules and the expression of hepatic genes, from hAECs. This work was supported by Wakayama Prefecture Collaboration of Regional Entities for the Advancement of Technological Excellence of the JST, and by a Grant-in-Aid for the 21st Century COE Program of the Japan MEXT.

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Electron microscopy analysis of degenerating pancreatic ß cells in transgenic mice expressing the MHC Class I antigen Dd

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100160297 ◽

1990 ◽

Vol 48 (3) ◽

pp. 548-549

Author(s):

T. A. Stewart ◽

D. Liggitt ◽

S. Pitts ◽

L. Martin ◽

M. Siegel ◽

...

Keyword(s):

Type I Diabetes ◽

Disease Process ◽

Class Ii ◽

Class I ◽

Type I ◽

Aberrant Expression ◽

Mhc Molecules ◽

Endogenous Insulin ◽

Class Ii Mhc ◽

Ifn Γ

Insulin-dependant (Type I) diabetes mellitus (IDDM) is a metabolic disorder resulting from the lack of endogenous insulin secretion. The disease is thought to result from the autoimmune mediated destruction of the insulin producing ß cells within the islets of Langerhans. The disease process is probably triggered by environmental agents, e.g. virus or chemical toxins on a background of genetic susceptibility associated with particular alleles within the major histocompatiblity complex (MHC). The relation between IDDM and the MHC locus has been reinforced by the demonstration of both class I and class II MHC proteins on the surface of ß cells from newly diagnosed patients as well as mounting evidence that IDDM has an autoimmune pathogenesis. In 1984, a series of observations were used to advance a hypothesis, in which it was suggested that aberrant expression of class II MHC molecules, perhaps induced by gamma-interferon (IFN γ) could present self antigens and initiate an autoimmune disease. We have tested some aspects of this model and demonstrated that expression of IFN γ by pancreatic ß cells can initiate an inflammatory destruction of both the islets and pancreas and does lead to IDDM.

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Expression of class I and class II MHC antigens in rat kidneys and definition of a distinct endothelial-monocyte syste,

Human Immunology ◽

10.1016/0198-8859(81)90139-7 ◽

1981 ◽

Vol 3 (4) ◽

pp. 380

Keyword(s):

Class Ii ◽

Class I ◽

Mhc Antigens ◽

Class Ii Mhc ◽

Class Ii Mhc Antigens ◽

Definition Of ◽

Rat Kidneys

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Role of Class-II Major Histocompatibility Complex (MHC)-Antigen–Positive Donor Leukocytes in Transfusion-Induced Alloimmunization to Donor Class-I MHC Antigens

Blood ◽

10.1182/blood.v92.2.690.414k12_690_694 ◽

1998 ◽

Vol 92 (2) ◽

pp. 690-694

Author(s):

K.J. Kao ◽

M.L.U. del Rosario

Keyword(s):

Immune Response ◽

Peripheral Blood ◽

Class Ii ◽

Class I ◽

Mononuclear Leukocytes ◽

Class I Mhc ◽

Mhc Antigens ◽

Mhc Molecules ◽

Class Ii Mhc ◽

Mhc Antigen

It has been shown that peripheral-blood mononuclear leukocytes (MNL) are responsible for transfusion-induced alloimmunization to donor major histocompatability complex (MHC) antigens. However, it is not known which subset of MNL is responsible for this immune response. Because elimination of class-II MHC antigen-positive passenger leukocytes effectively prolongs the survival of allografts, it has been hypothesized that class-II positive MNL are responsible for immunizing transfusion recipients to donor MHC antigens. To test this hypothesis, two different approaches were used. First, we compared the alloantigenicity of BALB/c mice (H-2d) peripheral blood MNL before and after depletion of class-II positive cells. CBA mice (H-2k) were used as transfusion recipients. Antibody development to donor class-I H-2 antigens was determined by flow cytometry and enzyme-linked immunoassay. After four weekly transfusions of MNL depleted for class-II positive cells, only 25% of recipient mice developed antibodies to donor H-2d antigens. In contrast, all mice transfused with control MNL became immunized. Second, we studied the alloantigenicity of peripheral MNL from C57BL/6 mice (H-2b) with homozygous deficiency of class-II MHC molecules in H-2 disparate recipient mice. After transfusions with class-II MHC molecule-deficient MNL, 0% of BALB/c, 40% of C57BR, and 25% of CBA-recipient mice developed antibodies to donor H-2b antigen. All control recipient mice were immunized. The antibody activities of the controls were also higher than those in the treatment group who became immunized. Thus, our study shows that class-II MHC antigen-positive MNL play a significant role in transfusion-induced alloimmunization to donor class-I MHC antigens. The results also support the hypothesis that direct antigen presentation by donor class-II positive MNL to the immune system of transfusion recipients is critical for the initiation of humoral immune response to donor MHC antigens.

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Expression of class I and class II MHC antigens in neuromuscular diseases

Journal of the Neurological Sciences ◽

10.1016/0022-510x(89)90023-3 ◽

1989 ◽

Vol 89 (2-3) ◽

pp. 213-226 ◽

Cited By ~ 74

Author(s):

Rhoda M. McDouall ◽

Michael J. Dunn ◽

Victor Dubowitz

Keyword(s):

Neuromuscular Diseases ◽

Class Ii ◽

Class I ◽

Mhc Antigens ◽

Class Ii Mhc ◽

Class Ii Mhc Antigens

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Role of Class-II Major Histocompatibility Complex (MHC)-Antigen–Positive Donor Leukocytes in Transfusion-Induced Alloimmunization to Donor Class-I MHC Antigens

Blood ◽

10.1182/blood.v92.2.690 ◽

1998 ◽

Vol 92 (2) ◽

pp. 690-694 ◽

Cited By ~ 16

Author(s):

K.J. Kao ◽

M.L.U. del Rosario

Keyword(s):

Immune Response ◽

Peripheral Blood ◽

Class Ii ◽

Class I ◽

Mononuclear Leukocytes ◽

Class I Mhc ◽

Mhc Antigens ◽

Mhc Molecules ◽

Class Ii Mhc ◽

Mhc Antigen

Abstract It has been shown that peripheral-blood mononuclear leukocytes (MNL) are responsible for transfusion-induced alloimmunization to donor major histocompatability complex (MHC) antigens. However, it is not known which subset of MNL is responsible for this immune response. Because elimination of class-II MHC antigen-positive passenger leukocytes effectively prolongs the survival of allografts, it has been hypothesized that class-II positive MNL are responsible for immunizing transfusion recipients to donor MHC antigens. To test this hypothesis, two different approaches were used. First, we compared the alloantigenicity of BALB/c mice (H-2d) peripheral blood MNL before and after depletion of class-II positive cells. CBA mice (H-2k) were used as transfusion recipients. Antibody development to donor class-I H-2 antigens was determined by flow cytometry and enzyme-linked immunoassay. After four weekly transfusions of MNL depleted for class-II positive cells, only 25% of recipient mice developed antibodies to donor H-2d antigens. In contrast, all mice transfused with control MNL became immunized. Second, we studied the alloantigenicity of peripheral MNL from C57BL/6 mice (H-2b) with homozygous deficiency of class-II MHC molecules in H-2 disparate recipient mice. After transfusions with class-II MHC molecule-deficient MNL, 0% of BALB/c, 40% of C57BR, and 25% of CBA-recipient mice developed antibodies to donor H-2b antigen. All control recipient mice were immunized. The antibody activities of the controls were also higher than those in the treatment group who became immunized. Thus, our study shows that class-II MHC antigen-positive MNL play a significant role in transfusion-induced alloimmunization to donor class-I MHC antigens. The results also support the hypothesis that direct antigen presentation by donor class-II positive MNL to the immune system of transfusion recipients is critical for the initiation of humoral immune response to donor MHC antigens.

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