Kato T, Honma K, Yamanaka A, Miura T, Okuda K
Oral Health Science Center, Department of Microbiology, Tokyo Dental College, 1-2-2 Masago, Mihama-ku, Chiba, Japan.
FEMS Immunol Med Microbiol. 2000 May;28(1):67-70. doi: 10.1111/j.1574-695X.2000.tb01458.x.
We investigated the heterogeneity of the humoral immune responses to whole cells and lipopolysaccharide (LPS) of Actinobacillus actinomycetemcomitans serotype b and production of cytokines in inbred strains of mice. Nine such strains were tested: A/J (H-2(a)), C57BL/6 (H-2(b)), BALB/c (H-2(d)), DBA/2 (H-2(d)), B10.BR (H-2(k)), C3H/He (H-2(k)), C3H/HeJ (H-2(k)), DBA/1 (H-2(q)) and B10.S (H-2(s)). Mice were immunized intraperitoneally with whole cells of A. actinomycetemcomitans ATCC 43718 (serotype b) in phosphate buffered saline (PBS; pH 7.2) emulsified with an equal volume of Freund's incomplete adjuvant. Serum immunoglobulin G (IgG), immunoglobulin A (IgA) and immunoglobulin M (IgM) levels against A. actinomycetemcomitans were measured by an ELISA system. ELISA analysis, using LPS fractions from serotype a, b or c strains of A. actinomycetemcomitans as the coating antigens, revealed that mice strains C3H/He, C3H/HeJ, B10.BR and B10.S had an extremely high-IgM response against serotype b LPS. High-IgM titer sera contain also elevated levels of IgA antibodies to the antigen. To compare the cytokine production among inbred mice, the amounts of interleukin-4 (IL-4), interleukin-5 (IL-5), and interleukin-6 (IL-6) released from mouse splenocytes were measured using ELISA systems specific for these cytokines. A. actinomycetemcomitans serotype b LPS stimulation induced IL-6 release from murine splenocytes of all tested strains. However, IL-4 and IL-5 were detected only in high-IgM/IgA responders to A. actinomycetemcomitans serotype b LPS, not in low-IgM/IgA responders. Thus, we found a relationship between the humoral immune response to LPS of A. actinomycetemcomitans serotype b and production of type 2 cytokines by splenocytes.
我们研究了近交系小鼠对伴放线放线杆菌b型血清型全细胞和脂多糖(LPS)的体液免疫反应的异质性以及细胞因子的产生。测试了9个这样的品系:A/J(H-2(a))、C57BL/6(H-2(b))、BALB/c(H-2(d))、DBA/2(H-2(d))、B10.BR(H-2(k))、C3H/He(H-2(k))、C3H/HeJ(H-2(k))、DBA/1(H-2(q))和B10.S(H-2(s))。将伴放线放线杆菌ATCC 43718(b型血清型)的全细胞与等体积的弗氏不完全佐剂乳化后,用磷酸盐缓冲盐水(PBS;pH 7.2)腹腔注射免疫小鼠。通过ELISA系统测定针对伴放线放线杆菌的血清免疫球蛋白G(IgG)、免疫球蛋白A(IgA)和免疫球蛋白M(IgM)水平。以伴放线放线杆菌a、b或c型血清型菌株的LPS组分作为包被抗原进行ELISA分析,结果显示C3H/He、C3H/HeJ、B10.BR和B10.S品系的小鼠对b型血清型LPS有极高的IgM反应。高IgM滴度血清中针对该抗原的IgA抗体水平也升高。为了比较近交系小鼠之间的细胞因子产生情况,使用针对这些细胞因子的ELISA系统测量从小鼠脾细胞释放的白细胞介素-4(IL-4)、白细胞介素-5(IL-5)和白细胞介素-6(IL-6)的量。伴放线放线杆菌b型血清型LPS刺激诱导了所有测试品系小鼠脾细胞释放IL-6。然而,仅在对伴放线放线杆菌b型血清型LPS有高IgM/IgA反应的小鼠中检测到IL-4和IL-5,在低IgM/IgA反应的小鼠中未检测到。因此,我们发现了伴放线放线杆菌b型血清型LPS的体液免疫反应与脾细胞产生2型细胞因子之间的关系。