Efficient recombination of a switch substrate retrovector in CD40-activated B lymphocytes: implications for the control of CH gene switch recombination.
Maturing B lymphocytes possess a recombination activity that switches the class of heavy chain Ig. The nature of the recombination activity, its molecular requirements and regulation remain elusive questions about B lymphocyte biology and development. Class switch recombination is controlled by cytokine response elements that are required to differentially activate CH gene transcription before their subsequent recombination. Here, we show that cultures of purified murine and human B cells, stimulated only by CD40 receptor engagement, possess a potent switch recombination activity. CD40 ligand-stimulated murine and human B lymphocytes were infected with recombinant retroviruses containing Smu and S gamma 2b sequences. Chromosomally integrated switch substrate retrovectors (SSRs), harboring constitutively transcribed S sequences, underwent extensive recombinations restricted to their S sequences with structural features akin to endogenous switching. SSR recombination commenced 4 days postinfection (5 days poststimulation) with extensive switch sequence recombination over the next 2 to 3 days. In contrast, endogenous S gamma 2b and S gamma 1 sequences did not undergo appreciable switch recombination upon CD40 signaling alone. As expected, IL-4 induced endogenous Smu to S gamma 1 switching, while endogenous Smu to S gamma 2b fusions remained undetectable. Surprisingly, IL-4 enhanced the onset of SSR recombination in CD40-stimulated murine B cells, with S-S products appearing only 2 days postinfection and reaching a maximum within 2 to 3 days. The efficiency of switch recombination with SSRs resembles that seen for endogenous C(H) class switching.
Department of Biochemistry and Cell Biology, Institute for Cell and Developmental Biology, State University of New York, Stony Brook 11794-5215, USA.
J
J
Ballantyne
D L
DL
Henry
J R
JR
Muller
F
F
Briere
C M
CM
Snapper
M
M
Kehry
K B
KB
Marcu
D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D019013:CD40 Antigens / Q000276:immunology* / Q000378:metabolism; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain / Q000187:drug effects; D005822:Genetic Vectors / Q000276:immunology*; D006801:Humans; D017578:Immunoglobulin Class Switching / Q000187:drug effects / Q000235:genetics*; D007127:Immunoglobulin Constant Regions / Q000235:genetics*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007142:Immunoglobulin gamma-Chains / Q000235:genetics; D007148:Immunoglobulin mu-Chains / Q000235:genetics; D015847:Interleukin-4 / Q000494:pharmacology; D007399:Interphase / Q000235:genetics / Q000276:immunology; D008213:Lymphocyte Activation / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008811:Mice, Inbred DBA; D014066:Palatine Tonsil / Q000276:immunology / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics*; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology; D013379:Substrate Specificity / Q000235:genetics
D019013:CD40 Antigens; D007127:Immunoglobulin Constant Regions; D007143:Immunoglobulin Heavy Chains; D007142:Immunoglobulin gamma-Chains; D007148:Immunoglobulin mu-Chains; D015847:Interleukin-4