Viable Borrelia burgdorferi enhances interleukin-10 production and suppresses activation of murine macrophages.
Although it is capable of eliciting strong innate and adaptive immune responses, Borrelia burgdorferi often evades immune clearance through largely unknown mechanisms. Our previous studies determined that infected interlukin-10-/- (IL-10-/-) mice show significantly lower B. burgdorferi levels than wild-type (B6) mice and that IL-10 inhibits innate immune responses critical for controlling B. burgdorferi infection. To determine whether virulent B. burgdorferi preferentially enhances IL-10 production, we developed an in vitro coculture medium (RPMI.B) in which both B. burgdorferi and primary macrophages (Mphis) remain viable. B. burgdorferi grew at similar rates and was able to regulate expression of immunoreactive proteins with similar kinetics in RPMI.B and in traditional BSK medium; in contrast, B. burgdorferi cultured in conventional tissue culture medium (RPMI) rapidly lost viability. Coculture of viable B. burgdorferi in RPMI.B with Mphis resulted in more rapid and significant increases in IL-10 transcripts and secreted proteins than coculture with nonviable B. burgdorferi in RPMI, which corresponded with decreased production of proinflammatory cytokines. Addition of live B. burgdorferi to Mphis in RPMI.B also elicited substantially higher IL-10 levels than heat-killed bacteria elicited, confirming that increased IL-10 production was not inherent to coculture in RPMI.B. Transfer of supernatants from B. burgdorferi-stimulated Mphis into naïve Mphi cultures resulted in suppressed activation upon subsequent stimulation with different bacterial agonists, and this suppression was obviated by IL-10-specific antibody. In vivo analyses determined that murine skin samples exhibited substantial upregulation of IL-10 within 24 h of injection of B. burgdorferi. Together, these results suggest that viable B. burgdorferi can suppress early Mphi responses during infection by causing increased release of IL-10.
Lazarus
John J
JJ
Department of Medical Microbiology and Immunology, University of Toledo Health Science Campus, 3000 Arlington Ave., Toledo, OH 43614, USA.
Kay
Maria A
MA
McCarter
Akisha L
AL
Wooten
R Mark
RM
D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis; D025065:Borrelia burgdorferi / Q000737:chemistry / Q000276:immunology*; D002478:Cells, Cultured; D018920:Coculture Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D020869:Gene Expression Profiling; D016753:Interleukin-10 / Q000096:biosynthesis* / Q000235:genetics; D008264:Macrophages / Q000737:chemistry / Q000276:immunology* / Q000382:microbiology*; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D050296:Microbial Viability; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000737:chemistry / Q000276:immunology / Q000382:microbiology; D015854:Up-Regulation
D001426:Bacterial Proteins; D008565:Membrane Proteins; D012333:RNA, Messenger; D016753:Interleukin-10