A new murine lymphocytotoxic monoclonal antibody recognizing HLA-A2, -A28 and -A9.
Monoclonal antibodies recognizing polymorphic as well as monomorphic epitopes on HLA antigens are important tools for understanding the immunobiology of HLA molecules. We immunized BALB/c mice with a HLA-A2 transfectant and screened for hybridomas which reacted with a HLA-A2 transfectant but not with a HLA-B75 transfectant. After subcloning by limiting dilution four times, a hybridoma secreting a monoclonal antibody (mAb) (IgG 2a, kappa) designated 1-145 was established. 1-145 reacted with Epstein-Barr virus transformed B lymphoblastoid cell lines (B cell lines) which expressed HLA-A2, -A28, -A23 and -A24. The titer of 1-145 in culture supernatant against HLA-A2 and -A28 antigens was similar and the titer against HLA-A23 was lower. 1-145 reacted with cells expressing HLA-A24 but the titer against HLA-A24 antigens was even lower than that against HLA-A23 antigens. The HLA-A24 antigens on the peripheral blood lymphocytes were not detected by 1-145 possibly due to the lower expression compared to the B cell lines. These differences of the titers were reflected to microlymphocytotoxicity assay in which 1-145 culture supernatant lysed all PBLs expressing HLA-A2,-A28 and -A23 but did not lyse PBLs expressing HLA-A24. Published deduced amino acid sequence data of HLA class 1 molecules indicate that Lys in position 127 may be critical for 1-145 binding.
Division of Research, Aichi Red Cross Blood Center, Japan.
S
S
Mizuno
F
F
Yako
H
H
Ohta
T
T
Kato
K
K
Wada
C
C
Uchigiri
M
M
Furuta
K
K
Ohya
T
T
Kurachi
T
T
Murase
T
T
Kamiya
K
K
Ozawa
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000937:Antigen-Antibody Reactions; D000961:Antilymphocyte Serum / Q000737:chemistry*; D015234:HLA-A Antigens / Q000276:immunology*; D015789:HLA-A2 Antigen / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data
D000911:Antibodies, Monoclonal; D000961:Antilymphocyte Serum; D015234:HLA-A Antigens; D015789:HLA-A2 Antigen; C037164:HLA-A28 antigen; C037709:HLA-A9 antigen