Effect of two antifilarials ivermectin (microfilaricidal) and CDRI Comp. 82/437 (macrofilaricidal) in combination with immunostimulator (CDRI Comp. 86/448) was evaluated on establishment of Acanthocheilonema viteae infection in Mastomys coucha. The immunostimulator along with the antifilarials was administered on single occasion (Day 0 of larval exposure). Immunostimulator when given in combination with macrofilaricidal agent (82/437), revealed significantly less percentage of worm recovery over untreated control as well as over treated (with either immunostimulator or antifilarial alone) infected controls. It is, thus surmised that establishment of filarial infection is affected by immunostimulant along with antifilarial agent

Bhatnagar Sunita

Chatterjee R.K.

Fatma Nigar

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T rop. Med., 37 (4), 151-157, December, 1995 151Acanthocheilonema viteae in Mastomys coucha:Combination Effect of An Immunostimulator (CDRI compound86/448) and Antifilarial agents on Establishment of InfectionSunita Bhatnagar, Nigar Fatma and R.K. ChatterjeeDivision of Parasitology, Central Drug Research InstituteLucknow 226 001, IndiaAbstract: Effect of two antifilarials ivermectin (microfilaricidal) and CDRI Comp. 82/437(macrofilaricidal) in combination with immunostimulator (CDRI Comp. 86/448) was evaluated onestablishment of Acanthocheilonema viteae infection in Mastomys coucha. The immunostimulatoralong with the antifilarials was administered on single occasion (Day 0 of larval exposure). Im-munostimulator when given in combination with macrofilaricidal agent (82/437), revealed sig-nificantly less percentage of wormrecovery over untreated control as well as over treated (witheither immunostimulator or antifilarial alone) infected controls. It is, thus surmised that estab-lishment of filarial infection is affected by immunostimulant along with antifilarial agent.Key words: Acanthocheilonema viteae, Mastomys coucha, Immunostimulator, Compound 82/437,Microfilaricidal, Parasite establishment, Macrofilaricidal, Compound 86/448.INTRODUCTIONFilariasis is a major parasitic disease of great public health importance affecting millions ofpeople in India alone (NICD, 1994). Apart from chemotherapeutic measures, control of the diseasein endemic areas largely depends upon the efficient preventive measures such as protection frominfective larval invasion from vector bites. It is now well documented that like many otherparasitic infections, filarial parasites also cause immunosuppression (Portaro et al, 1976; Ottesenet al, 1977; Piessens et al, 1980, a, b) which help them in their establishment in the host.Stimulation of host's immuneresponsiveness by use of immunomodulators thus appears to be alogical approach in counteracting establishment process of infective larvae. This has been amplysubstantiated from our earlier studies in which establishment of A. viteae and B. malayi infectionshas been shown to be jeopardized significantly when host was primed with immunomodulatorsbefore infective exposure (Misra et al, 1991; Murthy et al, 1992). On the other hand there arecertain antifilarial agents which are knownto act only partially on developing forms of filariae.Combination therapy with such antifilarial agents along with immunopotentiator appears to be apractical proposition in crusade against establishment of invading and developing larvae in thehost.Received for publication, December, 19, 1995CDRI Communication No. 5451152Previous studies in our laboratory have indicated that chemotherapeutic efficacy of an-tifilarial agent can also be enhanced by using it along with immunomodulators (Chatterjee et al,1988; Fatma et al, 1994). Thus, application of combination of immunostimulator and antifilarialagent might exert better and more efficient protection against infection. In the present study,efforts have been made to prevent the establishment of Acanthocheilonema viteae in Mastomyscoucha with combination therapy using CDRI compound 82/437 or ivermectin along with a syn-thetic immunostimulator, CDRI Compound 86/448.MATERIALS AND METHODSExperimental model: Acanthocheilonema viteae in Mastomys coucha was used as experimentalmodel. Six to eight weeks old male mastomys were exposed subcutaneously with 50 infectivelarvae (L3) of A. viteae collected from freshly dissected infected ticks (Ornithodoros moubata)following broadly the technique of Singh et al. (1988).Immunomodulator: CDRI Compound 86/448, a synthetic glycopeptide, was used in the presentstudy as immunostimulator. The Compound 86/448 is a structurally related congenor of muramyldipeptide (MDP) and was originally synthesized in this Institute (Haq et al, 1990).Antifilarials: Two antifilarials used in the study were ivermectin and a synthetic agent, com-pound 82/437. Ivermectin was procured from Merck Sharpe and Dohme, New Jersey. Thecompound 82/437 was originally synthesized in this Institute (Abuzar et al, 1986). The agent is abenzimidazole derivative (2,2' Dicarbomethoxy amino 5-5' dibenzimidazolyl ketone) havingmacrofilaricidal activity against three filarial species L. carinii, A. viteae and B. malayi in rodents(Fatma et al, 1989).Drugpreparation and Doseschedule: Compound 86/448 was prepared in sterile distilled waterand used at a dose of 250^g/animal on single occasion subcutaneously. Ivermectin and compound82/437 were used at 15^g/kg x 1 and 100mg/kg x 1 orally respectively. Both the compounds wereprepared in distilled water containing 0.1% Tween-80. Dose of antifilarials and immunomodu-lator corresponded to the ED50 values.Experiment: Four experimental groups, each consisting of 8- 10 animals were used. Differentgroups receiving treatment and controls were:1) treated with antifilarials only (ivermectin/Comp. 82/437).2) treated with immunomodulator only (86/448).3) treated simultaneously with antifilarial and immunomodulator.4) untreated controls.Animals belonging to different experimental groups were exposed to 50 L3 of A viteae 2 hrsafter administration of drug and/or immunomodulator. Untreated animals were also exposedunder identical conditions. The effect of individual or combination of drugs was assessed by ob-serving recovery of adult parasites on day 60 i. e., at the start of patency of infection.Adult wormrecovery: On day 60 of infective exposure, adult parasites were recovered fromsubcutaneous tissues of the infected animals. The percentage reduction in worm recovery wascalculated by comparing the number of live adult worms recovered from experimental animals153with that recovered from untreated infected controls.Statistical analysis: The data was analysed and p values were taken out using Student's T test.RESULTSEffect of Compounds 86/448 and ivermectin: Table 1 shows the effect of 86/448, ivermectin andtheir combination on establishment of A viteae infection in mastomys. Treatment with compound86/448 and ivermectin resulted in 60.2% reduction in adult worm recovery over control (p < 0.01).Ivermectin and compound 86/448 individually exerted respectively 52.8% and 42.9% reduction inwormrecovery over untreated infected control.Table 1. Effect of ivermectin, compound 82/437 and their combination with immunomodulator compound86/448 on establishment of A. viteae infection in M. couchaN°' of. No. of worms "÷T	5¢/uP”1"÷W,^^M^Uá"%̂reductionin %reductioninTreatment Dose* frná"f recovered *á"f worm wormrecoveryexts)» Mean±SD of worms recovery overcontrolI vermectin ( I ) 15//g/kgx 1 10 (3) 13.4±4.682/437 (1) lOOmg/kgx l 12 (3) 18.5±5.486/448 (1) 250//g/animalx l 12 (3) 16.2±4.4Ivermectin (F) 15j«g/kgx l 10 (3) ll.3±1.5887448 250^g/animal x l82/437 (V) lOOmg/kg x ! 12 (3)86/t448 25^/g/animal x 1I nfectedcontrol4 .3±3.8V ehicle 10 (3) 28.4+4.226.8±9.337.0±10.932.4±8.822.6±3.073.263.067.677.45 .7±7.£ 91.352.834.842.960.2 (P<0.01)84.8 (P<0.001)56.8±8.4 43.2 0.0C omp.86/448 was administered subcutaneously whereas other agents were fed orally*A11 animals were exposed to 50 L3 of A. viteaeTable la. Statistical significance of percent reduction in worm recovery(over control) amongst treated groups.T reated groups P valuesI VSIII VSMI VSFI VSVIvs1IvsFIvsV1vsF1vsVFvsVp<0.2p<0.5p<0.1p<0.005p<0.5p<0.001p<0.001p<0.02p<0.001p<0.001I =Ivermectin1 =82/4371 =86/448F =Ivermectin + 86/448V = 82/437 + 86/448154Effect of compounds 86/448 and 82/437: A significant reduction (84.8%) in establishment of A.viteae infection was observed when animals were exposed following treatment with combinationof compounds 86/448 and 82/437. Compound 82/437 alone exerted 34.8% reduction in adult wormrecovery over controls (Table 1). Statistical significance of percent reduction in wormrecovery(over control) amongst treated groups is shown in Table la.DISCUSSIONControl of fillariasis appears difficult in absence of any effective marcrofilaricide. Never-theless, continuous infective larval exposure through vectors in endemic area challenges ourstrategies in chemotherapy. One of the major logistics for control of filariasis would be the pro-tection of the population from establishment in host. This can be achieved either by vector controlor by drugs having lethal effect on developing forms of filarial larvae in the host. The latterstrategy i. e. control by use of filarial larvicidal agents has been evaluated in the present inves-tigation. Earlier, a number of attempts on prophylaxis by chemical agents have been made usingsingle or combination of antifilarial agents against different experimental filarial infections(Denham and Brandte, 1980; Zahner et al, 1987; McCall et al, 1993). Most of these studieshowever, did not yield desired results. The existing drug diethylcarbamazine (DEC) has strongeffect on microfilariae but its effect on other developing forms of target parasite, W.bancrofti isnot convincing (Jordan, 1958). However, the drug has been shown to be larvicidal with multipledoses as revealed in several experimental studies (Denham et al, 1978; Mak and Lim, 1983) in-cluding its use as chemoprophylactic agent against loaisis (Duke 1963).As immunesystem plays an important role in exertion of drug activity in host (Ottesen et.al, 1977; Kwaand Mak, 1980; Piessens et al, 1980) the strategy taken up in the present study isto boost the antifilarial larvicidal activity potential of the drug with the help of an im-munostimulator. Two candidate antifilarials CDRI compound 82/437 and ivermectin which havebeen shown earlier to exert moderate action against infective larvae (Fatma et al, 1989; Singh etal, 1990) have been used in the present investigation along with immunostimulator to potentiatetheir larvicidal property. It may be recalled that certain immunostimulators themselves can effectto an extent the growth, development and establishment of larvae in the host (Misra et al, 1991;Murthy et al, 1992).The single dose therapy with any individual antifilarial agent led to 35 to 53% effect onestablishment of adult wormswhen compared with controls. The maximumefficacy of 53% wasdemonstrated with ivermectin alone followed by immunostimulator compound 86/448 with 42.9%activity and an activity of 34.8% was displayed by the adulticidal agent, compound 82/437. It isinteresting to observe that though no significant enhancement in activity against wormestab-lishment occurred with combination of ivermectin and the immunostimulator, very significantimprovement in action of 82/437 occurred when this antifilarial was administered along withimmunostimulator. The better performance of combination of 82/437 and immunostimulator(86/448) was primarily due to complementary action of the two agents against establishment offilariids. In our earlier studies (Misra et. al, 1991) it was shown that peptide immunostimulators155like MDP and its derivatives could suppress filarial establishment in rodent hosts and subsequentdevelopment of microfilaraemia through enhancement in activity of reticuloendothelial systemincluding macrophage functioning. A. viteae used in the present study is known to be susceptibleto reactive oxygen intermediates (Batra et al, 1990). Nevertheless, the parasite also possesses anactive enzyme system like SOD and catalase to protect itself against host's oxidant attack.Compound 82/437 inhibits host's oxidant attack. Compound 82/437 also inhibits significantly theantioxidants (catalase and glutathione peroxidase) of parasite rendering them prone to H202toxicity leading to death (Batra et al, 1992). Thus on one hand presently used peptide im-munostimulator enhanced the release of reactive oxygen intermediates and on the other handcompound 82/437 disrupted the antioxidant defence system of the parasites leading to chain ofactivity against establishment of filarial larvae in the host.Though the immunostimulator used as combinant with ivermectin was the same, themechanism of exertion of activity of the two agents differ (Turner and Schaeffer, 1989; Mak et al,1991; Pal et al, 1991). None of these thus benefitted from the activity of the other with the resultthat no improvement in activity of the combination was observed. The combination of compound82/437 and the peptide immunostimulator should be investigated further for their efficacy againstestablishment of filarial infection in higher models eg. monkey with human filarial infection.ACKNOWLEDGEMENTSWeare thankful to Dr. K. B. Mathur, Division of Biopolymer, Central Drug Research In-stitute, Lucknow for kindly providing us immunomodulator used in the present study. Ourgrateful acknowledgements are also due to Indian Council of Medical Research and Council ofScientific and Industrial Research for financial support in the form of Research Associateshipsrespectively to SB and NF of us.REFERENCES1) Abuzar, S., Sharma, S., Fatma, N., Gupta, S., Murthy, P.K., Katiyar, J.C., Chatterjee, R.K. & Sen, A.B.(1986): Studies on potential filaricides Part 18: Synthesis of 2,2' dicarbomethoxyl amino 5-5' diben-zimidazolyl ketone and related compounds as potential anthelmintics. J. Med. Chem., 29, 1296 - 1299.2) Batra, S., Chatterjee, R.K. & Srivastava, V.M.L., (1990): Antioxidant enzymes inAcanthocheilonema viteaeand effect of antifilarial agents. Biochem. Pharmacol., 40, 2363 -2369.3) Batra, S., Chatterjee, R.K. & Srivastava, V.M.L. (1992): Antioxidant enzyme system in Litomosoidescarinii and Setaria cervi : Effect of a macrofilaricidal agent. Vet. Parasitol., 43, 93-103.4) Chatterjee, R.K., Fatma, N., Jain, R.K., Gupta, C.M. & Anand, N. (1988): Litomosoides carinii in rodents:Immunomodulators in potentiating action of diethylcarbamazine. Jpn. J. Exp. Med., 58, 243 -248.5) Denham, D.A. & Brandte, E. (1980): Chemoprophylactic activity of flubendazole against adult Brugiapahangi transplanted into the peritoneal cavities of jirds. J. Parasitol., 66, 933 -934.6) Denham, D.A., Suswillo, R.R., Rogers, R. & McGreevy, P.B. (1978): Studies with Brugia pahangi 17: Theanthelmintic effects of diethylcarbamazine. J. Parasitol., 64, 463 -468.7) Duke, B.O.L. (1963): Studies on the chemoprophylaxis of Loaisis It Observations on diethylcarbamazine156citrate (Banocide) as a prophylactic in man. Ann. Trop. Med. Parasitol., 57, 82-96.8) Fatma, N., Mathur, K.B. & Chatterjee, R.K. (1994): Chemotherapy of experimental filariasis: enhance-ment of activity profile of ivermectin with immunomodulators. Acta Tropica, 57, 55 - 67.9) Fatma, N., Sharnia, S. & Chatterjee, R.K. (1989): 2,2' Dicarbomethoxylamino 5,5' dibenzimidazolyl ke-tone a new antifilarial agent. Acta Tropica, 46, 311-321.10) Haq, W., Puri, A., Kundu, B., Saxena, R.P., Kapil, A.; Mathur, K.B. & Saxena, K.C. (1990): A processfor the synthesis of N-glycyl, N (L-abanyl-D-isoglutaminyl)-L-Cysyl-N-alkylamides possessing high im-munostimulant activity. Patent application filed (434/DEL/90).ll) Jordan, P. (1958): Action of DEC in vitro on infective larvae of W. bancrofti. Brit. J. Pharmacol.Chemother., 13, 318-320.12) Kwa, B.H. & Mak, J.W. (1980): Specific depression of cellular-mediated immunity in Malayan filariasis.Trans. R. Soc. Top. Med. Hyg., 74, 522.13) Mak, J.W. & Lim, P.K.C. (1983): Studies on the chemoprophylaxis of subperiodic Brugia malayi infectionin the leaf monkey (Preshytis melalophos) with diethylcarbamazine citrate. Z. Parasitenk., 69, 677-680.14) Mak, J.W., Navaratnam, V. & Ramachandran, C.P. (1991): Experimental Chemotherapy of lymphaticfilariasis. A review. Ann. Trop. Med. Parasitol., 85, 131- 137.15) McCall J.W., Coleman, S.U., Jones K.L., Petit, T.A. & Klei T.R. (1993): Prophylactic activity oftetracycline against B. pahangi infection in jirds (Meriones unguiculatus). J. Parasitol., 79, 775 - 776.16) Misra, S., Singh, D.P., Gupta, C.M., Chatterjee, R.K. & Anand, N. (1991): Acanthocheilonema viteae inMastomys natalensis : Effect of immunomodulator on establishment and course of infection. Med. Sci.Res., 19, 53-55.17) Murthy, P.K., Haq, W., Tyagi, K., Mathur, K.B. & Chatterjee, R.K. (1992): Effect of immunostimulatoron establishment and course of infection. Trop. Med., 34, 63-69.18) NICD (1994): Proceedings of National workshop to formulate strategy for Brugia malayi eradication fromIndia. 15 & 16th December NICD, Karaparamba, Calicut, Kerala, India.19) Ottesen, E.A., Walter, P.P. & Heck, L. (1977): Specific cellular unresponsiveness in human filariasis.Immunol., 33, 413-421.20) Pal, R., Anuradha, Rizvi, S.Y., Kundu, B., Mathur, K.B. & Katiyar, J.C. (1991): Leishmania donovani inhamster: Stimulation of non specific resistance by some novel glycopeptide and impact on therapeuticefficacy. Experientia, 47, 4010 -4016.21) Piessens, W.F., McGreevy, P.B., Piessens, P.W., McCreevy, M., Koiman, I., Saroso, J.S. & Dennis, D.T.(1980a): Immune response in human infections with Brugia malayi specific cellular responsiveness tofilarial antigens. J. Clin. Invest., 65, 172-179.22) Piessens, W.F., McGreevy, P.B., Ratiwayanto, S., McGreevy, M., Piessens, P.W., Koiman, L, Saroso,J.S. & Dennis, D.T. (1980b). Immune response in human infections with Brugia malayi correlation ofcellular and humoral reactions to microfilarial antigens with clinical status. Am. J. Trop. Med. Hyg., 29,563-570.23) Portaro, J.K., Britton, S. & Ash, L.R. (1976): Brugia pahangi : depressed mitogen reactivity in filarialinfections in the jirds (Meriones unguiculatus). Expt. Parasitol., 40, 438 -446.24) Singh, D.P., Misra, S., & Chatterjee, R.K. (1988): Response of Dipetalonema viteae in Mastomys natalensisto known antifilarials. Ind. J. Exp. Biol, 26, 48-52.25) Singh, D.P., Misra, S. & Chatterjee, R.K. (1990): A new technique in in vitro assay of antifilarials usingdifferent life forms of Acanthocheilonema viteae. Jap. J. Exp. Med., 60, 303-309.26) Turner, MJ. & Schaeffer, J.M. (1989): Eds. Campbell W.C. Mode of action of ivermectin. In. Ivermectin157& Abamectins Springer Verlag, New York, 73-88.27) Zahner, H., Striebel, H.P., Schultheiss, K., Schutze H.R. (1987): Chemoprophylactic activity of somebenzazole derivatives in experimental filarial infections. Trop. Med. Parasitol., 38, 259.

Acanthocheilonema viteae in Mastomys coucha : Combination Effect of An Immunostimulator (CDRI compound 86/448) and Antifilarial agents on Establishment of Infection

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Acanthocheilonema viteae in Mastomys coucha : Combination Effect of An Immunostimulator (CDRI compound 86/448) and Antifilarial agents on Establishment of Infection

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