The ID50 of wild-type was 5×103 spirochetes, whereas the ID50 of Δarp3 was 8×104 spirochetes. BTK inhibitor cost Relative infectivity could be restored by complementation of the Δarp3 mutant with lp28-1G, resulting in an ID50 identical to wild-type. Subsequent experiments in C3H and C3H-scid mice therefore used an infectious dose of 105 or greater spirochetes. Table 1 Dose-related infectivity of arp null (Δarp3), Δarp3-complemented (Δarp3 + lp28-1G) and wild-type B. burgdorferi in infant ICR mice, based upon culture of sub-inoculation site and urinary www.selleckchem.com/products/DMXAA(ASA404).html bladder at 2 weeks after inoculation Inoculum dose Δarp3 Δarp3 + lp28-1G wild-type 101

0/4* 0/4 0/4 102 0/4 0/4 0/4 103 0/4 0/4 0/4 104 1/4 4/4 4/4 105 2/4 4/4 4/4 * number of positive mice/number of mice tested. Four C3H-scid mice were each inoculated with 106 wild-type and five C3H-scid mice were each inoculated with 106 Δarp3 spirochetes,

and then necropsied at 60 days of infection to compare the full range of pathogenicity of each inoculum, unencumbered by acquired immunity. All inoculation sites and urinary bladders were culture-positive in both groups. Spirochetes were isolated from blood of 4/4 wild-type inoculated mice, whereas only 2/4 (one sample not collected) Δarp3 inoculated mice were bacteremic. All mice in both groups had severe (mean arthritis score 3.0 ± 0 SD) arthritis in tibiotarsal joints, as well as arthritis in both knees, and all mice had carditis. Despite equally severe disease, spirochete burdens in Selleckchem MRT67307 sub-inoculation, heart base, and tibiotarsal tissues, based upon flaB quantitative PCR (Q-PCR), were significantly

lower (P ≤ 0.05) in Δarp3 infected C3H-scid mice compared to wild-type infected mice (Figure 1). Spirochete burdens were also lower in ventricular muscle and quadriceps muscle, but differences were not statistically significant. Figure 1 Borrelia Carnitine palmitoyltransferase II burgdorferi flaB DNA copies per mg tissue weight (means ± standard deviations) in subinoculation site (subIN), heart base (HB), ventricular muscle (VM), quadriceps muscle (Quad) and tibiotarsus (Tibio) from 4 C3H- scid mice inoculated with wild-type (white bars) compared to 5 C3H- scid mice inoculated with arp null Δarp3 B. burgdorferi (black bars). (*, P ≤ 0.05). A confirmatory experiment was performed in which 5 C3H-scid mice were each inoculated with 106 wild-type and 5 C3H-scid mice were each inoculated with 106 Δarp3 spirochetes, and necropsied on day 28 after inoculation. Inoculation sites and urinary bladders in all mice from both groups were culture-positive, and all mice in both groups were bacteremic. Arthritis severity scores were equivalent in both groups (mean 2.8 ± 0.4 SD wild-type vs. mean 2.4 ± 0.5 SD Δarp3). Significantly lower flaB Q-PCR spirochete burdens (P ≤ 0.