975 - A Toll-Like Receptor 1 Polymorphism Is Associated with Heightened T Helper 1 Responses and Antibiotic-Refractory Lyme Arthritis

Monday, November 7, 2011: 9:00 AM-6:00 PM
Hall F2 - Poster Hall (McCormick Place West)
Klemen Strle, Junghee J. Shin, Lisa Glickstein and Allen C. Steere, Massachusetts General Hospital, Harvard Medical School, Boston, MA
Presentation Number: 975

Background/Purpose: Single nucleotide polymorphisms (SNPs) have been identified in several genes encoding Toll-like receptors (TLRs) that lead to modified cellular immune responses, decreased cytokine production, and altered disease susceptibility. The TLR SNPs with the best evidence for affecting immune function are TLR1 (1805GG), TLR2 (2258GA) and TLR5 (1174CT). We studied the frequency and functional outcome of these polymorphisms in patients with various manifestations of Lyme disease.

Method: PCR amplification-restriction fragment length polymorphism assays were used to determine the genotypes of TLR1 at position 1805, TLR2 at position 2258, and TLR5 at position 1174 in 71 patients with erythema migrans (EM) an early disease manifestation, and in 76 patients with antibiotic-responsive and 101 patients with antibiotic-refractory Lyme arthritis, a late disease manifestation. To assess the functional outcome of the polymorphisms, protein levels of 12 cytokines and chemokines were determined in serum of EM patients, and in joint fluid of Lyme arthritis patients using bead-based multiplex assays. In addition, the protein levels of these mediators were assessed in supernatants of Borrelia burgdorferi-stimulated PBMC from 43 patients with Lyme arthritis.

Result:The frequency of TLR1 polymorphism (1805GG) was greater in patients with antibiotic-refractory arthritis compared to patients with EM (62% vs, 49%, odds ratio = 1.7, P = 0.1) or antibiotic-responsive arthritis (62% vs. 47%, odds ratio = 1.9, P=0.05). Early in the illness, patients with EM who had 1805GG, primarily those infected with B. burgdorferi RST1 strains, had higher serum levels of IFNγ, CXCL9 and CXCL10, and more severe infection than patients with 1805TG/TT. These inflammatory responses were amplified in patients with Lyme arthritis, and the highest responses were observed in patients with antibiotic-refractory arthritis who had 1805GG and had been infected with RST1 strains. When PBMC from Lyme arthritis patients were stimulated with a B. burgdorferi RST1 strain, the 1805GG group had significantly larger fold changes in the levels of IFNγ, CCL2, CXCL9 and CXCL10, than those with 1805TG/TT (P<0.001). In contrast, the frequencies of TLR2 (2258GA) and TLR5 (1174CT) were not significantly different among the groups, and cytokine and chemokine values were similar in those with or without the polymorphism.

Conclusion: We concluded that the TLR1 1805GG polymorphism in B. burgdorferi RST1-infected patients was associated with stronger TH1-like immune responses, more symptomatic infection, and antibiotic-refractory arthritis. This is the first innate immune factor identified associated with antibiotic-refractory arthritis.

Keywords: Lyme disease, chemokines, cytokines, inflammation and polymorphism

Disclosure: K. Strle, None; J. J. Shin, None; L. Glickstein, None; A. C. Steere, None.