Direct measurement of lymphocyte receptor diversity.
The ability to mount an immune defense against infectious microorganisms and their products, and against tumors is believed to be a direct function of lymphocyte diversity. Because the diversity of lymphocyte receptor genes is >1000-fold more diverse than the entire genome and varies between genetically identical individuals, measuring lymphocyte diversity has been a daunting challenge. We developed a novel technique for measuring lymphocyte diversity directly using gene chips. We reasoned and here demonstrate that the frequency of hybridization of nucleic acids coding for lymphocyte receptors to the oligonucleotides on a gene chip varies in direct proportion to diversity. We applied the technique to detect changes in lymphocyte diversity in mice with known B cell alterations and in persons with known T cell repertoire defects. This approach is the first to provide direct analysis of lymphocyte receptor diversity and should facilitate fundamental study of the adaptive immune system and clinical efforts to assess immunological diseases. In addition, this approach could be more broadly applied, for example to measure diversity of viral quasi-species.
Ogle
Brenda M
BM
Transplantation Biology Program, Department of Physiology, Mayo Clinic, Rochester, Minnesota, MN 55905, USA.
Cascalho
Marilia
M
Joao
Cristina
C
Taylor
William
W
West
Lori J
LJ
Platt
Jeffrey L
JL
D000328:Adult; D000818:Animals; D000916:Antibody Diversity / Q000235:genetics; D015321:Gene Rearrangement / Q000235:genetics; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D019169:Jurkat Cells; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008875:Middle Aged; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D018075:RNA, Complementary / Q000235:genetics; D011946:Receptors, Antigen / Q000235:genetics*; D011947:Receptors, Antigen, B-Cell / Q000235:genetics; D011948:Receptors, Antigen, T-Cell / Q000235:genetics; D015203:Reproducibility of Results
D007143:Immunoglobulin Heavy Chains; D018075:RNA, Complementary; D011946:Receptors, Antigen; D011947:Receptors, Antigen, B-Cell; D011948:Receptors, Antigen, T-Cell; D012313:RNA