Recognition of major histocompatibility complex class II antigens by two anti-HLA-DR monoclonal antibodies on canine marrow cells correlates with effects on in vitro and in vivo hematopoiesis.
BACKGROUND
The role of major histocompatibility complex class II antigens in hematopoiesis is not well defined. We have shown that in vitro depletion of HLA-DR+ cells from canine marrow (e.g., by anti-HLA-DR monoclonal antibody [mAb] H81.9 and complement) prevents hematopoietic recovery. In vivo administration of the same mAb H81.9 after transplantation of unmanipulated autologous marrow results in graft failure. In vitro mAb H81.9 inhibited colony formation from short-term and long-term marrow cultures.


RESULTS
We investigated the effect of another mAb, Ca1.41, which also recognizes nonpolymorphic determinants on human (HLA-DR) and canine major histocompatibility complex class II antigens but is reactive with a narrower spectrum of cells in both canine peripheral blood and marrow than mAb H81.9 (and other anti-HLA-DR mAbs). In contrast to all other anti-HLA-DR mAbs tested, Ca1.41 did not interfere with colony formation in short-term or long-term marrow cultures and spared a population of small mononuclear cells with low forward light scatter that was eliminated via apoptosis by exposure to mAb H81.9. These target cells included lymphocytes and CD34+ hemopoietic precursors that expressed MHC class II molecules as determined by mAb H81.9 but not by mAb Ca1.41. In addition, transmembrane signaling and up-regulation of interleukin-1beta mRNA occurred with mAb H81.9 but not with Ca1.41. Transplantation of autologous marrow treated in vitro cytolytically with mAb Ca1.41 allowed for complete hematopoietic reconstitution. Further, in vivo administration of Ca1.41 posttransplant did not lead to autologous graft failure as had been observed with mAb H81.9.


CONCLUSIONS
These results support the notion that major histocompatibility complex class II is expressed on early hematopoietic precursor cells but recognition is dependent upon the mAb used. Preliminary studies show that mAb H81.9 triggered transmembrane signaling, resulting in up-regulation of interleukin-1beta and apoptosis, although mAb Ca1.41 did not. The fact that Ca1.41 binding was modified in the presence of exogenous invariant chain-derived peptide suggests that both binding and signaling are peptide dependent.
Yamaguchi
M
M
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA.
McSweeney
P A
PA
Kimball
L
L
Gersuk
G
G
Hong
D S
DS
Kwok
W
W
Storb
R
R
Beckham
C
C
Deeg
H J
HJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000494:pharmacology; D018952:Antigens, CD34 / Q000032:analysis; D017209:Apoptosis / Q000502:physiology; D001854:Bone Marrow Cells / Q000276:immunology* / Q000502:physiology*; D016026:Bone Marrow Transplantation; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D004285:Dogs; D005260:Female; D006684:HLA-DR Antigens / Q000276:immunology* / Q000502:physiology; D006410:Hematopoiesis / Q000187:drug effects / Q000502:physiology*; D000949:Histocompatibility Antigens Class II / Q000032:analysis / Q000276:immunology*; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D011184:Postoperative Period; D014182:Transplantation, Autologous
D000911:Antibodies, Monoclonal; D018952:Antigens, CD34; D016207:Cytokines; D006684:HLA-DR Antigens; D000949:Histocompatibility Antigens Class II