In Vitro Effects of Ivermectin and Sulphadiazine on Toxoplasma gondii

Abstract

Objective: Ivermectin and sulphadiazine were tested individually to determine their in vitro effects on Toxoplasma gondii grown in human epidermoid larynx carcinoma (Hep-2) cell culture. Study Design: In-vitro study. Material and Methods: Toxoplasma growth was quantities by an enzyme immunoassay performed directly on the fixed cultures, using a rabbit anti-T. gondii immunoglobulin G as the first antibody and a phosphatase-labeled anti-rabbit immunoglobulin G as the second antibody. For each drug, regression models were used to quantify the relationship between optical density values and antimicrobial agent concentrations in the cultures. Results: The 50% inhibitory concentrations (IC50) of ivermectin and sulphadiazine were found to be 0.2 μg/mL and 7.3 μg/mL after 48 h of exposure, respectively. None of the concentrations tested for each drugs demonstrated toxicity to Hep-2 cells after 72 h of incubation. Conclusion: These results indicate that ivermectin significantly inhibited replication of the tachyzoites of T. gondii RH strain. Turkish Başlık: İvermektin ve sulfadiazinin toxoplasma gondıı'ye etkinliklerinin in vitro değerlendirilmesi Anahtar Kelimeler: Toxoplasma gondii, ivermektin, sulfadiazin, in vitro, Hep-2 Amaç: İvermektin ile sulfadiazinin, in vitro T. gondii ile enfekte insan larenks epidermoid karsinom (Hep-2) hücrelerinde anti-toxoplasma aktiviteleri araştırılmıştır. Gereç ve Yöntemler: Antimikrobiyal ajanların anti-Toxoplasma aktiviteleri hücre kültür tabanlı ELISA ile değerlendirilmiştir. Primer antikor olarak tavşan anti-T gondii immunglobulin G sekonder antikor olarak alkalen fosfataz işaretli anti-rabbit immunglobulin G kullanılmıştır. Her ilaç için, optik dansite değerleri ile antimikrobiyal ajan konsantrasyonları arasındaki ilişki regresyon analizi uygulanarak değerlendirilmiştir. Bulgular: İvermektin ve sulfadiazinin inkübasyonun 48. saatinde, %50 inhibitör konsantrasyon (IC50) değerleri sırasıyla 0.2 μg/ml ve 7.3 μg/ml olarak hesaplanmıştır. Ayrıca her iki ilacın test edilen konsantrasyonlarının Hep-2 hücrelerine toksik olmadığı saptanmıştır. Sonuç: Bu sonuçlar; ivermektinin in vitro T.gondii RH suşu takizoit replikasyonunu anlamlı olarak inhibe ettiğini göstermiştir.

Keywords

• Toxoplasma gondii, ivermectin, sulphadiazine, in vitro, Hep-2

In Vitro Effects of Ivermectin and Sulphadiazine on Toxoplasma gondii

Öz

References

1. Montaya JG, Liesenfeld O. Toxoplasmosis. The Lancet 2004;363:1965-76. [CrossRef]
2. Montoya JG, Kovacs JA, Remington JS. Toxoplasma gondii. In:Mandell GL, Bennett JE, Dolin R, editors. Mandell, Douglas and Bennett’s Principles and Practice of Infectious Diseases. 6th ed. Philadelphia:Elsevier Press;2008. p.3170-92.
3. Wilson M, Jones LJ, McAuley LB. Toxoplasma. In:Murray PR, Baron EJ, Jorgensen JH, Landry ML, Pfaller MA, editors. Manual of Clinical Microbiology. 9th ed. Washington:ASM Press;2005. p.2070-81. Petersen E. Toxoplasmosis. Seminars in Fetal & Neonatal Medicine 2007;12:214-23. [CrossRef]
4. Merli A, Canessa A, Melioli G. Enzyme immunoassay for evaluation of Toxoplasma gondii growth in tissue culture. J Clin Microbiol 1985;21:88-91.
5. Derouin F, Chastang C. Enzyme Immunoassay to assess effect of antimicrobial agents on Toxoplasma gondii in tissue cultures. Antimicrob Agents Chemother 1988;32:303-7. [CrossRef]
6. Diab MR, El-Bahy MM. Toxoplasma gondii:Virulence of tachyzoites in serum free media at different temperatures. Exp Parasitol 2008;118:75-9. [CrossRef]
7. Meneceuri P, Bouldouyre MA, Aubert D, Villena I, Menotti J, Sauvage V, et al. Toxoplasma gondii:In vitro susceptibility of various genotypic strains to pyrimethamine, sulphadiazine and atovaquone. Antimicrob Agents Chemother 2008;52:1269-77. [CrossRef]
8. Derouin F, Chastang C. In vitro effects of folate inhibitors on Toxoplasma gondii. Antimicrob Agents Chemother 1989;33:1753-9. [CrossRef]

9. Guzzo CA, Furtek CI, Chen C, Tipping R, Clineschmidt CM, Sciberras DG, et al. Safety, tolerability, pharmacokinetics of escalating high doses of ivermectin in healthy adult subjects. J Clin Pharmacol 2002;42:1122-33. [CrossRef] 600.00 500 500.00 400 400.00 300 300.00