Reconstitution of functional human B lymphocytes in NOD/SCID mice engrafted with ex vivo expanded CD34(+) cord blood cells.
OBJECTIVE
Functional capacity of B cells developed from ex vivo expanded hematopoietic stem cells has not been fully evaluated. Therefore, we investigated the antigen-specific antibody production in human B cells maturated from ex vivo expanded cord blood (CB) CD34(+) cells in NOD/Shi-scid (NOD/SCID) mice.


METHODS
CB CD34(+) cells were cultured for 5 days in the presence of human cytokines and the murine stromal cell line HESS-5, and transplanted into irradiated NOD/SCID mice. These mice, reconstituted with human hematopoietic cells, were challenged with T-cell-independent (TI) or T-cell-dependent (TD) antigens after CD19(+) cells appeared at 6 weeks.


RESULTS
Three months later, anti-dinitrophenol (DNP)-specific antibody was detected in both mice immunized with DNP-Ficoll (TI) and those immunized with DNP-keyhole limpet hemocyanin or DNP-ovalbumin (TD). The anti-DNP antibody was mainly immunoglobulin M, but a small amount of immunoglobulin G also was detected. In the spleen, the majority of CD19(+) cells expressed mature B-cell markers such as CD40, immunoglobulin M, immunoglobulin D, cytoplasmic Cmu, and light chains kappa, and lambda.


CONCLUSIONS
These results indicate that human B cells develop from CD34(+) cells in NOD/SCID mice to produce antigen-specific antibody with in vivo primary stimulation. This system provides a powerful and versatile tool for studying the entire process of human B-lymphocyte development and producing specific human monoclonal antibodies.
Li
Chenwgwen
C
Department of Hematology, Tokai University School of Medicine, Kanagawa, Japan.
Ando
Kiyoshi
K
Kametani
Yoshie
Y
Oki
Masayuki
M
Hagihara
Masao
M
Shimamura
Kazuo
K
Habu
Sonoko
S
Kato
Shunichi
S
Hotta
Tomomitsu
T
D000818:Animals; D000918:Antibody Specificity; D018941:Antigens, CD19 / Q000032:analysis; D018952:Antigens, CD34 / Q000032:analysis; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology; D001854:Bone Marrow Cells / Q000502:physiology; D019013:CD40 Antigens / Q000032:analysis; D019070:Cell Lineage; D002478:Cells, Cultured / Q000187:drug effects / Q000637:transplantation; D018920:Coculture Techniques; D004136:Dinitrobenzenes / Q000276:immunology; D005312:Fetal Blood / Q000166:cytology*; D005856:Germ-Free Life; D006412:Hematopoietic Stem Cells / Q000187:drug effects; D006801:Humans; D007114:Immunization; D007074:Immunoglobulin G / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D007231:Infant, Newborn; D008213:Lymphocyte Activation; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D011828:Radiation Chimera / Q000097:blood / Q000276:immunology; D013154:Spleen / Q000166:cytology; D033581:Stem Cell Transplantation; D017154:Stromal Cells / Q000502:physiology; D014183:Transplantation, Heterologous
D018941:Antigens, CD19; D018952:Antigens, CD34; D019013:CD40 Antigens; D004136:Dinitrobenzenes; D007074:Immunoglobulin G; D007075:Immunoglobulin M