Analysis of main T-cell subsets and activated T supressor/cytotoxic cells in patients with Borrelia burgdorferi s. lato only infection and co-infections with Anaplasma phagocytophilum, Bartonella spp. and Babesia microti

Department of Infectious Diseases, Medical University of Lublin, Poland
Department of Allergology and Environmental Hazards, Institute of Rural Health, Lublin, Poland
Ann Agric Environ Med 2016;23(1):111–115
The study was designed to assess the role of some important immunologic factors with regards to both laboratory results and clinical symptoms in patients with confirmed Lyme disease. Additional examinations were carried out for co-infections with a number of tick-borne pathogens.

Material and Methods:
The study group consisted of 54 patients with Lyme disease and a group of 21 healthy controls. Serology of co-infections with Anaplasma phagocytophilum, Bartonella spp. and Babesia microti was carrieed out in all patients. Blood samples were stained using the whole-blood lysis method and analyzed concurrently on a flow cytometer FACSCalibur. Directly conjugated anti-human monoclonal antibodies against CD3, CD4, CD8, CD16, CD56, HLA-DR and CD69 were used.

No significant differences were observed with respect to thepretreatment level of CD4+ and CD8+ cells. In patients with symptoms relief and symptoms persistence, lower percentages of CD4+ and CD8+ cells were found, but with no statistical dependence. In the study group, both in patients with and without co-infections, pretreatment values of CD16+CD56+ cells did not differ significantly. In patients who did not respond to the treatment, the baseline percentage of NK cells was higher (P<0.01) than in group with clinical improvement, and lower after the treatment, whereas in patients with symptoms relief after the treatment there was an increase in the percentage of NK cells.

Co-infections with Anaplasma phagocytophilum, Bartonella spp. and Babesia microti had no impact on T-cell percentages in Lyme disease patients. There was a lower baseline percentage of NK cells in patients not responding to antibiotic treatment.

Krzysztof Tomasiewicz   
Department of Infectious Diseases, Medical University of Lublin, Poland
1. Auwaerter PG Point: antibiotic therapy is not the answer for patients with persisting symptoms attributable to lyme disease. Clin Infect Dis. 2007; 45: 143–148.
2. Sjovall J. Clinical and immunological aspects of Lyme boreliosis. Linkoping 2011.
3. Swanson SJ, Neitzel D, Reed KD, Belongia EA. Coinfections acquired from Ixodes ticks. Clin Microbiol Rev. 2006; 19: 708–727.
4. Smith T, Kilbourne FL. Investigations into the nature, causation and prevention of Texas or southern cattle fever. Bur Anim Ind Bull. 1983; 1: 151–152.
5. Herwaldt B, Persing DH, Precigout EA, Goff WL, Mathiesen DA, et al. A fatal case of babesiosis in Missouri: identification of another piroplasm that infects humans. Ann Intern Med. 1996; 124: 643–650.
6. Rios L, Alvarez G, and Blair S. Serological and parasitological study and report of the first case of human babesiosis in Colombia . Rev Soc Bras Med Trop. 2003; 36: 493–498.
7. Bakken J S, Dumler JS, Chen SM, Eckman MR, Van Etta LL, et al. Human granulocytic ehrlichiosis in the upper Midwest United States. A new species emerging? JAMA 1994; 272: 212–218.
8. Tomasiewicz K, Modrzewska R, Buczek A, Stanczak J, Maciukajc J. The risk of exposure to Anaplasma phagocytophilum infection in Mid-Eastern Poland. Ann Agric Environ Med. 2004; 11: 261–264.
9. Telford III SR, Wormser GP. Bartonella spp. transmission by ticks not established. Emerg Infect Dis. 2010;16: 379–384.
10. Billeter SA, Levy MG, Chronel BB, Breitschwerdt EB. Vector transmission of Bartonella species with emphasis on the potential for tick transmission. Med Vet Entomol. 2008; 22: 1–15.
11. Billeter SA, Miller MK, Breitschwerdt EB, Levy MG. Detection of two Bartonella tamiae–like sequences in Amblyomma americanum (Acari: Ixodidae) using 16S–23S intergenic spacer region–specific primers. J Med Entomol. 2008; 45: 176–9.
12. Sjöwall J, Fryland L, Nordberg M, Sjögren F, Garpmo U, et al. Decreased Th1-type inflammatory cytokine expression in the skin is associated with persisting symptoms after treatment of erythema migrans. PLoS ONE 2011; 6: e18220.
13. Lubega J. T-helper 1 versus T-helper 2 lymphocyte immunodysregulation is the central factor in genesis of Burkitt lymphoma: hypothesis. Infect Agent Cancer. 2007; 2: 1230–1234.
14. Singh R, Haghjoo E, Liu F. Cytomegalovirus M43 gene modulates T helper cell response. Immunology Letters. 2003; 88: 31–35.
15. Logar M, Ruzić-Sabljić E, Maraspin V, Lotrić-Furlan S, Cimperman J, et al. Comparison of erythema migrans caused by Borrelia afzelii and Borrelia garinii Infection 2004; 32: 15–19.
16. Dong Z, Edelstein M, Glickstein L. CD8+ T cells are activated during the early Th1 and Th2 immune responses in a murine Lyme disease model. Infect Immun. 1997; 65: 5334–5337.
17. Gross DM, Steere AC, Huber BT. T helper 1 response is dominant and localized to the synovial fluid in patients with Lyme arthritis. J Immunol. 1998; 160: 1022–1028.
18. Moro MH, Bjornsson J, Marietta EV, Hofmeister EK, Germer JJ, et al. Gestational attenuation of Lyme arthritis is mediated by progesterone and IL-4. J. Immunol 2001; 166: 7404–7409.
19. Ganapamo F, Dennis V A, Philipp MT. Differential acquired immune responsiveness to bacterial lipoproteins in Lyme disease-resistant and -susceptible mouse strains. Eur J Immunol. 2003; 33: 1934–1940.
20. Steere AC, Taylor E, McHugh GL, Logigian EL. The overdiagnosis of Lyme disease. JAMA 1993; 269: 1812–1816.
21. Zeinder NS, Dolan MC, Massung R, Piesman J, Fish D. Coinfection with Borrelia burgdorferi and the agent of human granulocytic ehrlichiosis suppresses IL-2 and IFN γ production and promotes an IL-4 response in C3H/HeJ mice. Parasite Immunol. 2000; 22: 581–588.
22. Moro M H, Zegarra-Moro OL, Bjornsson J, Hofmeister EK, Bruinsma E, et al. Increased arthritis severity in mice coinfected with Borrelia burgdorferi and Babesia microti. J Infect Dis. 2002; 186: 428–431.
23. Coleman JL, Levine D, Thill C, Kuhlow C, Benach JL . Babesia microti and Borrelia burgdorferi follow independent courses of infection in mice. J Infect Dis. 2005; 192: 1634–1641.
24. Krause PJ, McKay K, Thompson CA, Sikand VK, Lentz R, et al. Disease-specific diagnosis of coinfecting tickborne zoonoses: babesiosis, human granulocytic ehrlichiosis, and Lyme disease. Clin Infect Dis. 2002; 34: 1184–1111.
25. Krause PJ, Telford III SR, Spielman A, Sikand V, Ryan R, et al. Concurrent Lyme disease and babesiosis. Evidence for increased severity and duration of illness. JAMA 1996; 275: 1657–1660.
26. Stricker RB, Winger EE. Decreased CD57 lymphocyte subset in patients with chronic Lyme disease. Immunol Lett 2001; 76: 43–48.
27. Marques A, Brown MR, Fleisher TA. Natural killer cell counts are not different between patients with post-Lyme disease syndrome and controls. Clin Vaccine Immunol. 2009; 16: 1249–1250.
28. Sticker RB. Natural Killer Cells in Chronic Lyme Disease. Clin Vaccine Immunol. 2009; 16: 1704–1706.
29. Mihaylova I, DeRuyter M, Rummens JL, Bosmans E, Maes M. Decreased expression of CD69 in chronic fatigue syndrome in relation to inflammatory markers: evidence for a severe disorder in the early activation of T lymphocytes and natural killer cells. Neuro Endocrinol Lett. 2007; 28: 477–483.