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HLA antigens in newly established four choriocarcinoma cell lines
P 83 CHARACTERIZATION OF MAJOR HISTOCOMPATIBILITY COMPLEX (MHC) ANTIGEN EXPRESSION BY FIRST TRIMESTER CHORIONIC VILLI
J.A. LATA, J.B. SMITH, R.S. TUAN, M.M. M U L L I G A N A N D L.G. J A C K S O N , Departments of Medicine, Orthopaedic Surgery and Genetics, Thomas Jefferson University, Philadelphia,P A 19107 USA Our studies of antigen expression of human chorionic villi have b e e n conducted on tissue obtained by chorionic villus sampling (CVS) at 9-10 weeks of gestation. We have examined expression of MHC antigens and t h e i r corresponding mRNAs by i m m u n o h i s t o c h e m i s t r y and in situ cDNA-mRNA hybridization. Distinct regions of the s3rncytiotrophoblast have b e e n found to express an HLA Class I monomorphic d e t e r m i n a n t as d e t e c t e d by the mouse monoclonal antibody V16/32. These areas are also stained by trophoblast specific antibodies including antibodies to c y t o k e r a t i n and human p l a c e n t a l lactogen. DR expression was not d e t e c t e d on m e s e n c h y m e or on cell layers immunohistologically designated as trophoblast. In situ cDNA-mRNA hybridization studies were conducted on p a r a f f i n sections of villi employing cDNA probes for HLA-B7 and DR chains to localize mRNA encoding Class I and II d e t e r m i n a n t s . Hybridization of Class I cDNA was evident in both the m e s e n c h y m e and trophoblast layers of the villi. By contrast, no signal was evident for DR transcripts. Investigations of mechanisms involved in the regulation of MHC antigen expression are currently underway.
M H C ; chorionic villus, m R N A
P 84 HLA A N T I G E N S
IN N E W L Y E S T A B L I S H E D FOUR C H O R I O C A R C I N O M A CELL LINES
SETSUKO GOTO, M I T S U R U SAIT0, T O M O M I T S U 0KAMOTO, Y O H K 0 NISHIKAWA, OSAMU YAMAMURO, A K I H I R O NAWA, YASUSHI N A G A T A A N D Y U T A K A T O M O D A . Dept of Obstet & Gyneco, N a g o y a U n i v e r s i t y School of medicine, 65 T s u r u m a l - c h o , Showa-ku, Nagoya 466, Japan Objects We e x a m i n e d HLA antigens in the four c h o r i o c a r c i n o m a cell lines, those were newly e s t a b l i s h e d in our l a b o r a t o r y and n a m e d as NaUCC-I, -2, -3, and -4 respectively. Methods Immunoa d h e r e n c e assay and p r o t e i n - A assay were p e r f o r m e d to detect HLA a n t i g e n s on those cell lines. Furthermore, u s i n g HLA-B7 cDNA and HLA-A2 genomic DNA, S o u t h e r n b l o t a n a l y s i s w a s p e r f o r m e d a b o u t t o t a l D N A s e x t r a c t e d f r o m t h o s e c h o r i o c a r c i n o m a cells. Results In s e r o l o g i c a l analysis, NaUCC-I, -2, -3, and -4 were reacted to the a n t i b o d i e s against both m o n o m o r p h i e d e t e r m i n a n t s of HLA class 1 a n t i g e n s and b e t a 2 - m i c r o g l o b u l l n . A n t i b o d i e s to p o l y m o r p h l c d e t e r m i n a n t s not only of the p a t i e n t ' s HLA type but a l s o of the h u s b a n d ' s type reacted to NaUCC-I and -2. S o u t h e r n blot a n a l y s i s r e v e a l e d that all cell lines h a d gene w h i c h c o d e d c l a s s 1 HLA, a n d that the l e n g t h p o l y m o r p h i s m existed in the EcoRI DNA fragment of all the cell lines by h y b r i d i z i n g w l t h H L A - A 2 g e n o m l c DNA.
Keywords:
choriocarcinoma,
HLA,
S o u t h e r n blot.
81
J.A. LATA, J.B. SMITH, R.S. TUAN, M.M. M U L L I G A N A N D L.G. J A C K S O N , Departments of Medicine, Orthopaedic Surgery and Genetics, Thomas Jefferson University, Philadelphia,P A 19107 USA Our studies of antigen expression of human chorionic villi have b e e n conducted on tissue obtained by chorionic villus sampling (CVS) at 9-10 weeks of gestation. We have examined expression of MHC antigens and t h e i r corresponding mRNAs by i m m u n o h i s t o c h e m i s t r y and in situ cDNA-mRNA hybridization. Distinct regions of the s3rncytiotrophoblast have b e e n found to express an HLA Class I monomorphic d e t e r m i n a n t as d e t e c t e d by the mouse monoclonal antibody V16/32. These areas are also stained by trophoblast specific antibodies including antibodies to c y t o k e r a t i n and human p l a c e n t a l lactogen. DR expression was not d e t e c t e d on m e s e n c h y m e or on cell layers immunohistologically designated as trophoblast. In situ cDNA-mRNA hybridization studies were conducted on p a r a f f i n sections of villi employing cDNA probes for HLA-B7 and DR chains to localize mRNA encoding Class I and II d e t e r m i n a n t s . Hybridization of Class I cDNA was evident in both the m e s e n c h y m e and trophoblast layers of the villi. By contrast, no signal was evident for DR transcripts. Investigations of mechanisms involved in the regulation of MHC antigen expression are currently underway.
M H C ; chorionic villus, m R N A
P 84 HLA A N T I G E N S
IN N E W L Y E S T A B L I S H E D FOUR C H O R I O C A R C I N O M A CELL LINES
SETSUKO GOTO, M I T S U R U SAIT0, T O M O M I T S U 0KAMOTO, Y O H K 0 NISHIKAWA, OSAMU YAMAMURO, A K I H I R O NAWA, YASUSHI N A G A T A A N D Y U T A K A T O M O D A . Dept of Obstet & Gyneco, N a g o y a U n i v e r s i t y School of medicine, 65 T s u r u m a l - c h o , Showa-ku, Nagoya 466, Japan Objects We e x a m i n e d HLA antigens in the four c h o r i o c a r c i n o m a cell lines, those were newly e s t a b l i s h e d in our l a b o r a t o r y and n a m e d as NaUCC-I, -2, -3, and -4 respectively. Methods Immunoa d h e r e n c e assay and p r o t e i n - A assay were p e r f o r m e d to detect HLA a n t i g e n s on those cell lines. Furthermore, u s i n g HLA-B7 cDNA and HLA-A2 genomic DNA, S o u t h e r n b l o t a n a l y s i s w a s p e r f o r m e d a b o u t t o t a l D N A s e x t r a c t e d f r o m t h o s e c h o r i o c a r c i n o m a cells. Results In s e r o l o g i c a l analysis, NaUCC-I, -2, -3, and -4 were reacted to the a n t i b o d i e s against both m o n o m o r p h i e d e t e r m i n a n t s of HLA class 1 a n t i g e n s and b e t a 2 - m i c r o g l o b u l l n . A n t i b o d i e s to p o l y m o r p h l c d e t e r m i n a n t s not only of the p a t i e n t ' s HLA type but a l s o of the h u s b a n d ' s type reacted to NaUCC-I and -2. S o u t h e r n blot a n a l y s i s r e v e a l e d that all cell lines h a d gene w h i c h c o d e d c l a s s 1 HLA, a n d that the l e n g t h p o l y m o r p h i s m existed in the EcoRI DNA fragment of all the cell lines by h y b r i d i z i n g w l t h H L A - A 2 g e n o m l c DNA.
Keywords:
choriocarcinoma,
HLA,
S o u t h e r n blot.
81