Alloantibodies to PHA-Activated Lymphocytes Detect Human Qa-Like Antigens

Gazit, Ephraim; Gothelf, Yaël; Gil, R.; Orgad, Shlomit; Pitman, Theodore; Watson, Ada; Yang, Soo Young; Yunis, Edmond J.

doi:10.1007/978-3-642-69770-8_156

Cited by 14 publications

(10 citation statements)

References 15 publications

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“…They assumed that the antigens detected by this reagent were analogous to Qa molecules. The additional P2m-associated molecules described here, which are expressed at high levels on activated cells, may be analogous to those described by Gazit et al (1984).…”

Section: Discussionsupporting

confidence: 63%

Biochemical characterization of activation‐associated bovine class I major histocompatibility complex antigens

Bensaïd¹,

Kaushal²,

MacHugh³

et al. 1989

Animal Genetics

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Utilizing a 'sandwich' ELISA assay we have been able to demonstrate that mAb W6/32, B1G6 and IL-A19 are reactive with three different monomorphic determinants on bovine class I major histocompatibility complex (MHC) molecules.Sequential immunoprecipitations performed with the mAb revealed that class I molecules on PBM comprise a single population with respect to reactivity with the mAb in that the P2m-associated proteins bear all three epitopes. By contrast, TCGF-driven lymphoblasts and cells transformed by Theileria parva (Tp) additionally express molecules of Mr 45000 bound to P2m which are recognized by mAb B1G6 and IL-A19 but not by W6/32. These two subclasses of molecules were further distinguished on the basis that, when tunicamycin was added to cultures in the preparation of cells for analysis, mAb W6/32 precipitated class I heavy chains of Mr 39000 while the extra molecules detected only by mAb B1G6 and IL-A19 were of Mr 37000 and 39000. On thymocytes, the mAb W6/32-non-reactive class I molecules are present in low amounts and are expressed by cells in the medulla area, unlike BoTl (analogous to human CD1) molecules which are expressed by the cortical cells. Our studies also revealed that the supposed P2m-specific mAb B1G6 does not recognize the P2m-associated molecules (BoT1) precipitated by mAb TH97A and thus the specificity of mAb B1G6 in cattle is for an epitope on bovine p2m which is strongly influenced by the nature of the heavy chain with which the P2m is associated. 241 242 A. Bensaid et al. heterozygous animal (Bensaid et al. 1988). Here we describe studies utilizing mAb IL-A19, detecting a monomorphic epitope of BOLA class I proteins, and mAb TH97A, defining BoTl molecules analogous to human CD1 (MacHugh et al. 1988).We have compared, with histo-and immunochemical techniques, the properties of the molecules carrying the determinants recognized by these mAb with those of the molecules carrying determinants for the cross-reacting mAb W6/32 and B1G6 in order to gain further insights into the diversity, structure and expression of class I MHC molecules in cattle. Materials and methods AnimalsA 3-month-old Boran (Bos indicus) calf was used as a donor of thymus cells. Peripheral blood mononuclear cells were collected from a 4-year-old Boran steer. Monoclonal antibodiesMAb W6/32 (IgG2a) recognizes an epitope on all HLA-A, B and C heavy chains associated with p2m (Barnstable et al. Parham et al. 1979). The mAb B1G6 (IgG2b) is directed against human P2m and recognizes this molecule both free and complexed with HLA heavy chains (Liabeuf etal. 1981). B1G6 has also been found to be reactive with bovine P2m (Chardon et al. 1983).The mAb IL-A19 (Ig2a) was derived in our laboratory from a hybridoma produced by fusion of the spleen cells of a Balb/c mouse with myeloma X63 .Ag. Prior to fusion, the mouse had received multiple inoculations with PBM from B. indicus and B. tuurus animals. Hybridoma selection and cloning was by standard methods. This mAb reacts with an epitope present on the surface of PBM of all ...

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Section: Discussionsupporting

confidence: 63%

Biochemical characterization of activation‐associated bovine class I major histocompatibility complex antigens

Bensaïd¹,

Kaushal²,

MacHugh³

et al. 1989

Animal Genetics

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“…The medium was also supplemented with 10% of either PHS or FCS. As the source for IG2, we used locally prepared IG2 (Gazit et al 1984) or recombinant IL-2. 30% of the local I L 2 or 5 unitdm1 of the recombinant IL2 were used.…”

Section: Methodsmentioning

confidence: 99%

“…Two types of PHA blasts were used: F-blasts prepared according to the method of Fauchet et al (Fauchet et al 1986) and G-blasts prepared in Leiden according to the method of Gazit et al (Gazit et al 1984).…”

Section: Cellsmentioning

confidence: 99%

The Leiden “Class IV” or “Class I‐like” workshop November 20–23, 1986

Leeuwen¹,

D'Amaro²,

Fauchet³

et al. 1988

Tissue Antigens

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“…In contrast, occasional multiparous sera contain antibodies reactive with nonclassical antigens expressed on activated T cells that are analogous to murine Qa antigens. [93][94][95][96][97][98][99][100][101][102] In our laboratory, we found that hybridization of polyrA) + RNA from embryonic and trophoblast cells taken from four individuals with the synthetic DNA probes common total HLA-family gene and with the HLA-A,-B,-C locus-specific synthetic DNA probes revealed a single component 18S species, characteristic of class I transcripts and with almost equal levels of expression in both tissues. On the other hand, Northern blot analysis employing the locus-specific synthetic probes revealed that a very low amount of hybridization was observed in trophoblast RNA, while the 18S species was readily seen in embryonic cell RNA from all four individuals.…”

Section: Novel Versus Classical Mhc-antigens Expressed On Trophoblastmentioning

confidence: 99%

EDITORIAL: Is Production of Blocking Antibodies in Successful Human Pregnancy an Epiphenomenon?

Takeuchi

1990

American J Rep Immunol

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Alloantibodies to PHA-Activated Lymphocytes Detect Human Qa-Like Antigens

Cited by 14 publications

References 15 publications

Biochemical characterization of activation‐associated bovine class I major histocompatibility complex antigens

Biochemical characterization of activation‐associated bovine class I major histocompatibility complex antigens

The Leiden “Class IV” or “Class I‐like” workshop November 20–23, 1986

EDITORIAL: Is Production of Blocking Antibodies in Successful Human Pregnancy an Epiphenomenon?

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