HLA-G, -E, -F preworkshop: tools and protocols for analysis of non-classical class I genes transcription and protein expression.
Non-classical MHC class I HLA-E, -F, and -G molecules differ from classical class I histocompatibility antigens by specific patterns of transcription, protein expression, and immunological functions. Restriction of the expression pattern of these non-classical antigens may play a key role in modulation of immune responses during pregnancy and diseases but remains to be additionally defined. A specific component of the second International Conference on HLA-G and the 13th HLA-G Histocompatibility Workshop will be dedicated to the analysis of transcription and expression of non-classical class I genes in normal and pathological tissues. In a first step, referred to as the preworkshop, we here report the analysis and conclusions of a working group which was constituted to gather and validate optimal reagents and protocols allowing RT-PCR analysis of HLA-E, -F, -G transcript levels and flow cytometry and immunochemistry analysis of HLA-G expression in cells and tissues. As a result of this work, use of specific primers and probes detecting alternative transcripts of HLA-E, -F, and G have been validated in transfected cells expressing differential pattern of HLA class I antigens. Analysis of the specificity and affinity of collected antibodies has allowed definition of reagents to be proposed for immunochemistry and flow cytometry analysis of HLA-G expression in normal and pathological tissues during the workshop. This work has allowed constitution of an extended workshop group which is now initiating analysis of non-classical class I transcription and expression in various cells and tissues, a collective contribution that will additionally refine our view of the expression of these antigens in normal and pathological situations.
Paul
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Service de Recherches en Hémato-Immunologie, CEA/DRM/DSV, Hôpital St Louis, Institut Universitaire d'Hématologie, 1 Avenue Claude Vellefaux, 75475, Cedex 10, Paris, France. paul@dsvidf.cea.fr
Rouas-Freiss
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N
Moreau
P
P
Cabestre
F A
FA
Menier
C
C
Khalil-Daher
I
I
Pangault
C
C
Onno
M
M
Fauchet
R
R
Martinez-Laso
J
J
Morales
P
P
Villena
A A
AA
Giacomini
P
P
Natali
P G
PG
Frumento
G
G
Ferrara
G B
GB
McMaster
M
M
Fisher
S
S
Schust
D
D
Ferrone
S
S
Dausset
J
J
Geraghty
D
D
Carosella
E D
ED
D000911:Antibodies, Monoclonal / Q000276:immunology; D002460:Cell Line; D005434:Flow Cytometry / Q000379:methods*; D015870:Gene Expression; D005805:Genes, MHC Class I; D006680:HLA Antigens / Q000235:genetics / Q000378:metabolism; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D014162:Transfection; D000097625:HLA-E Antigens
D000911:Antibodies, Monoclonal; D006680:HLA Antigens; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I; D000097625:HLA-E Antigens; C062341:HLA-F antigens