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Infection and Immunity, May 2003, p. 2938-2940, Vol. 71, No. 5
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.5.2938-2940.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Humoral Immunity Reflects Altered T Helper Cell Bias in Borrelia burgdorferi-Infected T-Cell-Deficient Mice

Linda K. Bockenstedt,^1* Marie-Claude Shanafelt,¹ Alexia Belperron,¹ Jialing Mao,¹ and Stephen W. Barthold²

Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06520,¹ Center for Comparative Medicine, University of California, Davis, California 95616²

Received 26 November 2002/ Returned for modification 30 December 2002/ Accepted 14 January 2003

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In murine Lyme borreliosis, the absence of T lymphocytes augments the T helper cell 2 type humoral response but does not alter disease susceptibility. Arthropod transmission of Borrelia burgdorferi spirochetes results in similar antibody isotypes when T cells are present, suggesting that vector effects can negate T-cell functions in vivo.

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Lyme borreliosis is a multisystem illness caused by infection with the arthropod-borne spirochete Borrelia burgdorferi (7, 27). In the mouse model of Lyme borreliosis, spirochetes introduced intradermally by syringe inoculation or by the tick vector first establish infection in the skin (2, 26), where they may encounter T lymphocytes among the responding immune cells. T cells have been shown to influence the T helper (Th) cell response and alter resistance to disease due to intracellular pathogens (10, 11, 13, 16-18), but their effects on the immune response to a vector-borne extracellular pathogen are unknown. We therefore used mice that had been rendered deficient in T cells either by antibody (Ab) depletion (BALB/c mice) or by targeted disruption of the T-cell receptor (TCR) gene (B6.129 TCR ^-/- mice [B6.129P2-Tcrd^tm1Mom; Jackson Laboratories, Bar Harbor, Maine] and BALB/c TCR ^-/- mice [29], a gift of Mark Shlomchik, Yale University) to examine the consequences of T-cell deficiency on the course of murine Lyme borreliosis. For Ab depletion, 150 µg of purified hamster anti-mouse TCR monoclonal Ab UC7-13D5 or 150 µl of hamster serum was subcutaneously injected into 4- to 5-week-old BALB/c mice daily, beginning 5 days prior to experimental infection and continuing for a total of 9 days.

Mice were infected by either intradermal inoculation of 10⁴ cloned N40 spirochetes in 100 µl of Barbour Stoenner Kelly medium (1) or by infestation with five N40-infected Ixodes scapularis nymphs (5). Fourteen days after infection, lymph node cells from T-cell-deficient BALB/c mice produced less gamma interferon (IFN-) than controls, as measured by cytokine-specific enzyme-linked immunosorbent assay (ELISA) (Fig. 1). Interleukin 4 was detected in culture supernatants of stimulated lymph node cells, consistent with the known Th2 dominance of BALB/c mice (4, 19), and the level was modestly increased in the absence of T cells (273 versus 408 pg/ml; P < 0.07). A more dramatic reduction in IFN- production was noted for B6.129 TCR ^-/- mice (Fig. 1), but no interleukin 4 was detected. Thus, the absence of T cells in B. burgdorferi infection reduces the Th1 cytokine response, consistent with previous studies showing that T cells can direct polarization of ß Th-cell subsets (11, 17, 18).

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FIG. 1. Lymph node cells from mice deficient in T cells produce less IFN-. Production of IFN- by popliteal lymph node cells isolated from individual mice was measured by ELISA. Each result is reported as the mean of values obtained for mice within each group ± the standard error of the mean.

ELISA measurement of B. burgdorferi-specific immunoglobulin G (IgG) isotypes (25) reflected the reduction in the Th1 cytokine IFN- (Table 1). BALB/c TCR ^-/- mice had a significant rise in endpoint titers of IgG1 (P = 0.0317, Mann- Whitney test) and tended to have lower IgG2a levels. In contrast, B6.129 TCR ^-/- mice exhibited a marked reduction in IgG3 endpoint titers (P = 0.0159, Mann-Whitney test), and titers of IgG2b trended higher. One explanation for the different IgG isotypes induced in the two mouse strains is that T cells serve to decrease the genetically dominant Th2 response in BALB/c mice, whereas they promote a Th1 response in B6.129 mice.

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TABLE 1. Anti-B. burgdorferi IgG and IgG isotype reciprocal endopoint ELISA titersa

Saliva from I. scapularis nymphs, the vector for B. burgdorferi, contains factors that inhibit host defenses and retard the development of immunologic resistance to tick infestation (21, 22, 24). Although deposited locally in the skin during tick feeding, tick saliva appears to have systemic effects. The feeding of I. scapularis ticks on mice results in decreased IFN- production and downregulation of a Th1-type response by mitogen-stimulated splenocytes (23, 28). Interestingly, in our study, B. burgdorferi-specific IgG isotypes induced in control mice by vector-borne infection paralleled those seen in TCR ^-/- mice infected with cultured spirochetes, supporting a vector-induced bias toward Th2 cytokine patterns. Tick-transmitted spirochetes elicited predominantly IgG1 in wild-type BALB/c mice, at levels comparable to those achieved in BALB/c TCR ^-/- infected with cultured spirochetes (Table 1). The absence of T cells did not lead to a further increase in this Ab subset after vector-borne infection.

Despite the altered cytokine and humoral immune responses in B. burgdorferi-infected T-cell-deficient mice, we observed no difference in the severity or prevalence of disease as assessed by histopathology (3). When examined using TCR ^-/- mice on two backgrounds or after depletion of T cells by monoclonal Ab treatment, no significant differences were noted in arthritis or carditis (Table 2). The relative decrease in B. burgdorferi-specific Th1 responses in TCR ^-/- mice did not lead to protracted episodes of carditis even though CD4⁺ Th1 cells have been shown to promote resolution of this disease manifestation (6). Two previous studies also failed to show an effect of absence of the Th1 cytokine IFN- on murine Lyme arthritis (8, 14). Mean pathogen burden among mouse groups was the same as assessed by quantitative PCR of the spirochete ospA gene in urinary bladders (data not shown).

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TABLE 2. Absence of T cells does not alter disease in B. burgdorferi-infected micea

B. burgdorferi TCR ^-/- mice were able to develop protective Ab. Both wild-type and TCR ^-/- mice that were passively immunized with 500 µl of 1:5 dilution of immune serum from 45-day-infected TCR ^-/- mice 24 h prior to challenge infection with tick-borne spirochetes were completely protected as assessed by culture and histopathology (data not shown) (3). This was an expected outcome, because for B. burgdorferi infection, protective and arthritis-resolving antibodies arise in the absence of T-cell help (12, 20).

In summary, we used the murine model of Lyme borreliosis to investigate the effect of T cells on the adaptive immune response to a vector-borne extracellular pathogen. While disease expression was not altered, our results show that T cells influence the quality of the humoral immune response to B. burgdorferi introduced through the skin and suggest that tick transmission of spirochetes negates the T-cell effect. T-cell effector functions have been implicated in a variety of inflammatory and infectious processes (9, 15), yet the degree to which these cells play a part in the host immune response remains uncertain. In mammals, the location of these cells at sites of anatomic barriers to the environment suggests a primary role in the early immune response against agents, including arthropod vectors, which can penetrate the barrier. Given the similarity between IgG isotypes induced by vector-borne infection in wild-type mice to those of TCR ^-/- mice, it is possible that the Th cell bias attributed to tick saliva may be due to its actions on dendritic epidermal T cells. These cells may contribute to the local cytokine milieu for maturing dendritic cells in the skin and influence the priming of /ß T-cell responses in the lymph nodes. Although T-cell effects do not alter the outcome from B. burgdorferi infection, our results have important implications for other vector-borne pathogens transmitted through the skin.

 
We thank Debbie Beck for excellent technical assistance and Ruth Montgomery for editorial review of the manuscript.

This work was supported by NIH AR42637, AI50604, and AR47058 and the Arthritis Foundation (L.K.B.) and NIH AI26815 (S.W.B.).

 
* Corresponding author. Mailing address: 609 LCI, Section of Rheumatology, Yale University School of Medicine, 333 Cedar St., New Haven, CT 06520-8031. Phone: (203) 785-7063. Fax: (203) 785-7053. E-mail: linda.bockenstedt{at}yale.edu.

Editor: F. C. Fang

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1
1. Barthold, S. W., M. S. de Souza, J. L. Janotka, A. L. Smith, and D. H. Persing. 1993. Chronic Lyme borreliosis in the laboratory mouse. Am. J. Pathol. 143:419-420.[Abstract]
2
2. Barthold, S. W., D. H. Persing, A. L. Armstrong, and R. A. Peeples. 1991. Kinetics of Borrelia burgdorferi dissemination and evolution of disease following intradermal inoculation of mice. Am. J. Pathol. 139:263-273.[Abstract]
3
3. Barthold, S. W., C. L. Sidman, and A. L. Smith. 1992. Lyme borreliosis in genetically resistant and susceptible mice with severe combined immunodeficiency. Am. J. Trop. Med. Hyg. 47:605-613.
4
4. Bix, M., Z.-E. Wang, B. Thiel, N. J. Schork, and R. M. Locksley. 1998. Genetic regulation of commitment to interleukin 4 production by a CD4+ T cell-intrinsic mechanism. J. Exp. Med. 188:2289-2299.[Abstract/Free Full Text]
5
5. Bockenstedt, L. K., E. Hodzic, S. Feng, K. W. Bourrel, A. deSilva, R. R. Montgomery, E. Fikrig, J. D. Radolf, and S. W. Barthold. 1997. Borrelia burgdorferi strain-specific Osp C-mediated immunity in mice. Infect. Immun. 65:4661-4667.[Abstract]
6
6. Bockenstedt, L. K., I. Kang, C. Chang, D. Persing, A. Hayday, and S. W. Barthold. 2001. CD4+ T helper 1 cells facilitate regression of murine Lyme carditis. Infect. Immun. 69:5264-5269.[Abstract/Free Full Text]
7
7. Bockenstedt, L. K., and S. E. Malawista. 1995. Lyme disease, p. 1234-1249. In R. R. Rich (ed.), Clinical immunology. Mosby-Year Book, St. Louis, Mo.
8
8. Brown, C. R., and S. L. Reiner. 1999. Experimental Lyme arthritis in the absence of interleukin-4 or gamma interferon. Infect. Immun. 67:3329-3333.[Abstract/Free Full Text]
9
9. Carding, S. R., and P. J. Egan. 2002. T cells: functional plasticity and heterogeneity. Nat. Rev. Immunol. 2:336-345.[CrossRef][Medline]
10
10. Cardona, A. E., and J. M. Teale. 2002. / T cell-deficient mice exhibit reduced disease severity and decreased inflammatory response in the brain in murine neurocysticercosis. J. Immunol. 169:3163-3171.[Abstract/Free Full Text]
11
11. Ferrick, D. A., M. D. Schrenzel, T. Mulvania, B. Hsieh, W. G. Ferlin, and H. Lepper. 1995. Differential production of interferon-gamma and interleukin-4 in response to Th1- and Th2-stimulating pathogens by gamma delta T cells in vivo. Nature 373:255-257.[CrossRef][Medline]
12
12. Fikrig, E., S. W. Barthold, M. Chen, I. S. Grewal, J. Craft, and R. A. Flavell. 1996. Protective antibodies in murine Lyme disease arise independently of CD40 ligand. J. Immunol. 157:1-3.[Abstract]
13
13. Fu, Y.-X., C. E. Roark, K. Kelly, D. Drevets, P. Campbell, R. O'Brien, and W. Born. 1994. Immune protection and control of inflammatory tissue necrosis by T cells. J. Immunol. 153:3101-3115.[Abstract]
14
14. Glickstein, L., M. Edelstein, and J. Z. Dong. 2001. Gamma interferon is not required for arthritis resistance in the murine Lyme disease model. Infect. Immun. 69:3737-3743.[Abstract/Free Full Text]
15
15. Hayday, A. C. 2000. cells: a right time and a right place for a conserved third way of protection. Annu. Rev. Immunol. 18:975-1026.[CrossRef][Medline]
16
16. Hiromatsu, K., Y. Yoshikai, G. Matsuzaki, S. Ohga, K. Muramori, K. Matsumoto, J. A. Bluestone, and K. Nomoto. 1992. A protective role of / T cells in primary infection with Listeria monocytogenes in mice. J. Exp. Med. 175:49-56.[Abstract/Free Full Text]
17
17. Huber, S., C. Shi, and R. C. Budd. 2002. Gamma delta T cells promote a Th1 response during coxsackievirus B3 infection in vivo: role of Fas and Fas ligand. J. Virol. 76:6487-6494.[Abstract/Free Full Text]
18
18. Huber, S. A., D. Graveline, W. K. Born, and R. L. O'Brien. 2001. Cytokine production by Vgamma(+)-T-cell subsets is an important factor determining CD4(+)-Th-cell phenotype and susceptibility of BALB/c mice to coxsackievirus B3-induced myocarditis. J. Virol. 75:5860-5869.[Abstract/Free Full Text]
19
19. Kang, I., S. W. Barthold, D. H. Persing, and L. K. Bockenstedt. 1997. T helper cell cytokines in the early evolution of murine Lyme arthritis. Infect. Immun. 65:3107-3111.[Abstract]
20
20. McKisic, M. D., and S. W. Barthold. 2000. T-cell-independent responses to Borrelia burgdorferi are critical for protective immunity and resolution of Lyme disease. Infect. Immun. 68:5190-5197.[Abstract/Free Full Text]
21
21. Nuttall, P. A., G. C. Paesen, C. H. Lawrie, and H. Wang. 2000. Vector-host interactions in disease transmission. J. Mol. Microbiol. Biotechnol. 2:381-386.[CrossRef][Medline]
22
22. Ribeiro, J. M. C., G. T. Makoul, J. Levine, D. R. Robinson, and A. R. Spielman. 1985. Antihemostatic, antiinflammatory, and immunosuppressive properties of the saliva of a tick. Ixodes dammini. J. Exp. Med. 161:332-344.[Abstract/Free Full Text]
23
23. Schoeler, G. B., S. A. Manweiler, and S. K. Wikel. 1999. Ixodes scapularis: effects of repeated infestations with pathogen-free nymphs on macrophage and T lymphocyte cytokine responses of BALB/c and C3H/HeN mice. Exp. Pathol. 92:239-248.
24
24. Schoeler, G. B., and S. K. Wikel. 2001. Modulation of host immunity by haematophagous arthropods. Ann. Trop. Med. Parasitol. 95:755-771.[CrossRef][Medline]
25
25. Shanafelt, M.-C., I. Kang, S. W. Barthold, and L. K. Bockenstedt. 1998. Modulation of murine Lyme borreliosis by interruption of the B7/CD28 T cell costimulatory pathway. Infect. Immun. 66:266-271.[Abstract/Free Full Text]
26
26. Shih, C.-M., R. J. Pollack, S. R. Telford, III, and A. Spielman. 1992. Delayed dissemination of Lyme disease spirochetes from the site of deposition in the skin of mice. J. Infect. Dis. 166:827-831.[Medline]
27
27. Steere, A. C. 2001. Lyme disease. N. Engl. J. Med. 345:115-124.[Free Full Text]
28
28. Zeidner, N., M. L. Mbow, M. Dolan, R. Massung, E. Baca, and J. Piesman. 1997. Effects of Ixodes scapularis and Borrelia burgdorferi on modulation of the host immune response: induction of a Th2 cytokine response in Lyme disease-susceptible (C3H/HeJ) mice but not in disease-resistant (BALB/c) mice. Infect. Immun. 65:3100-3106.[Abstract]
29
29. Zuany-Amorim, C., C. Ruffie, S. Haile, B. B. Vargaftig, P. Pereira, and M. Pretolani. 1998. Requirement for T cells in allergic airway inflammation. Science 280:1265-1267.[Abstract/Free Full Text]

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Infection and Immunity, May 2003, p. 2938-2940, Vol. 71, No. 5
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.5.2938-2940.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Bockenstedt, L. K. Articles by Barthold, S. W. Search for Related Content PubMed PubMed Citation Articles by Bockenstedt, L. K. Articles by Barthold, S. W.