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1.
Figure 4

Figure 4. CD28- and ICOS-dependent costimulation is critical for the induction of the secondary, but not primary, IgG anti-MCPS response to M1883. From: The nature of an in vivo anti-capsular polysaccharide response is markedly influenced by the composition and/or architecture of the bacterial subcapsular domain.

C57BL/6 (“Wild-type”), CD28−/− (C57BL/6 background), and ICOS−/− (C57BL/6 background) mice (7 per group) were immunized i.p. with 2 × 108 CFU of heat-inactivated intact M1883 in saline and boosted on day 21. Serum titers of Ag-specific IgG were determined by ELISA. Significance (*) p≤0.05 between “Wild-type” versus CD28−/− or ICOS−/− mice.

Swadhinya Arjunaraja, et al. J Immunol. ;188(2):569-577.
2.
Figure 2

Figure 2. The MCPS-specific IgM and IgG isotype responses to isolated MCPS and intact M1883 are distinct. From: The nature of an in vivo anti-capsular polysaccharide response is markedly influenced by the composition and/or architecture of the bacterial subcapsular domain.

BALB/c mice (7 per group) were immunized i.p. with 2 × 108 CFU of heat- inactivated intact M1883 or purified MCPS in saline, and boosted i.p. with the same dose on day 21. Serum titers of MCPS-specific IgM, IgG, and IgG subclasses (IgG3, IgG1, IgG2b and IgG2a) were determined. Significance (*) p≤0.05 between MenC and MCPS.

Swadhinya Arjunaraja, et al. J Immunol. ;188(2):569-577.
3.
Figure 7

Figure 7. Co-immunization of MenC and Pn14 promotes secondary boosting of the IgG anti-PPS14 response to Pn14. From: The nature of an in vivo anti-capsular polysaccharide response is markedly influenced by the composition and/or architecture of the bacterial subcapsular domain.

BALB/c mice (7 per group) were immunized i.p. with 2 × 108 CFU of heat-inactivated intact MenC ([A] strain M1883 or [B] strains FAM18 C+ or FAM18 C−]) or 2 × 108 CFU of heat-inactivated intact Pn14 (strain R614) or a combination of MenC and R614 (2 × 108 CFU each), and boosted in a similar fashion on day 21. Serum titers of Ag-specific IgG were determined by ELISA. Significance (*) p≤0.05 between mice immunized with single bacteria versus combination.

Swadhinya Arjunaraja, et al. J Immunol. ;188(2):569-577.
4.
Figure 5

Figure 5. CD40 ligand-dependent costimulation is critical for the induction of the secondary, but not primary, IgG anti-MCPS response to M1883. From: The nature of an in vivo anti-capsular polysaccharide response is markedly influenced by the composition and/or architecture of the bacterial subcapsular domain.

BALB/c mice (7 per group) were injected i.p. with either anti-CD40L mAb (clone MR1) or polyclonal hamster IgG (“Control”) [0.3 mg per mouse]. Mice were then immunized, 1 day later, i.p. with 2 × 108 CFU of heat- inactivated intact M1883 in saline and boosted 21 days later. Serum titers of Ag-specific IgG were determined by ELISA. Significance (*) p≤0.05 between “Control” versus anti-CD40L-injected mice.

Swadhinya Arjunaraja, et al. J Immunol. ;188(2):569-577.
5.
Figure 6

Figure 6. Endogenous TLR4, but not TLR2 or MyD88, signaling is critical for induction of peak primary and secondary serum titers of MCPS-specific IgG in response to M1883, but is not critical for secondary boosting. From: The nature of an in vivo anti-capsular polysaccharide response is markedly influenced by the composition and/or architecture of the bacterial subcapsular domain.

(A) C3H/HeJ(Tlr4Lps-d) and C3H/HeOuJ (control mice), or (B) TLR2−/− or (C) MyD88−/− mice (same group of control C57BL//6 mice used for both “B” and “C”) [7 per group] were immunized i.p. with 2 × 108 CFU of heat-inactivated intact M1883 in saline and boosted on day 21 and 35. Serum titers of MCPS-specific IgM, IgG, and IgG isotypes were determined by ELISA from sera obtained on the indicated days.

Swadhinya Arjunaraja, et al. J Immunol. ;188(2):569-577.
6.
Figure 1

Figure 1. The primary and secondary IgG anti-MCPS responses to intact MenC are similar to that elicited by a meningococcal conjugate vaccine. From: The nature of an in vivo anti-capsular polysaccharide response is markedly influenced by the composition and/or architecture of the bacterial subcapsular domain.

(A). BALB/c mice (7 per group) were immunized i.p. with 2 × 108 CFU of U.V.-inactivated intact MenC (strain M1883) in saline or 1µg of MCPS-TT in alum + CpG-ODN, and boosted i.p. with the same dose on day 28. Serum titers of Ag-specific IgM and IgG were measured by ELISA. Significance (*) p≤0.05 between secondary titers relative to peak primary titers (B). BALB/c mice (7 per group) were immunized i.p. with 2 × 108 CFU of heat-inactivated intact FAM18 C+ (encapsulated MenC) or FAM18 C− (unencapsulated MenC), and boosted i.p. with the same dose on day 21. Serum titers of Ag-specific IgG were determined by ELISA. Significance (*) p≤0.05 between FAM18 C+ and FAM18 C−.

Swadhinya Arjunaraja, et al. J Immunol. ;188(2):569-577.
7.
Figure 3

Figure 3. CD4+ T cells are required for the induction of the secondary, but not primary IgG anti-MCPS response to intact M1883. From: The nature of an in vivo anti-capsular polysaccharide response is markedly influenced by the composition and/or architecture of the bacterial subcapsular domain.

BALB/c mice (7 per group) were injected i.p. either with a depleting anti-CD4+ mAb (clone GK.1.5) or polyclonal rat IgG (“Control”) [0.5 mg per mouse]. Both Ab-injected mice and athymic nude mice (BALB/c background) were then immunized, 1 day later, i.p. with 2 × 108 CFU of heat-inactivated intact M1883 in saline, and boosted 21 days later in the absence of Ab. Serum titers of Ag-specific IgG were measured by ELISA. Significance (*) p≤0.05 between “Control” versus and-CD4 mAb-injected mice or athymic nude mice.

Swadhinya Arjunaraja, et al. J Immunol. ;188(2):569-577.

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