Immunocontraceptive potential of a recombinant chimeric protein composed by a single gonadotropin-releasing hormone molecule and B subunit of Escherichia coli heat-labile enterotoxin in a mice model
Lívia Budziarek Eslabão, Neida Lucia Conrad, Pedro Machado Medeiros de Albuquerque, Rodrigo Casquero Cunha, Renan Eugênio Araujo Piraine, Carine Dahl Corcini, Fábio Pereira Leivas Leite
Abstract
The present study aimed to evaluate the immunocontraceptive potential of a chimera containing gonadotropin-releasing hormone (GnRH) and the B subunit of Escherichia coli heat-labile enterotoxin (LTB) as a carrier molecule in male BALB/c mice. Mice were randomly divided into six experimental groups, according to the antigen/control inoculation: GnRH/LTB with adjuvant; commercial vaccine; GnRH/LTB; LTB with adjuvant; LTB; and PBS as control group. The humoral immune response was evaluated by indirect ELISA and the contraceptive potential was assessed through histological analysis of the animals’ gonads and serum testosterone. Vaccinated groups respond with production of anti-GnRH/LTB antibodies. GnRH/LTB associated with adjuvant induced significant (P < 0.05) higher IgG titer after 14 days after the prime vaccination, keeping high titer up to 126 days. The histology results showed that chimeric antigen caused changes in spermatogenesis, and in the gonads tissue. In addition, this group had significantly lower testosterone levels than the control group (P < 0.05). The chimera produced through the fusion of a single GnRH molecule with LTB as a carrier are capable of inducing the humoral immune response, blocking endocrine functions related to reproduction, and causing tissue alterations in the gonads of male mice.
References
Aucouturier, J., Dupuis, L., & Ganne, V. (2001). Adjuvants designed for veterinary and human vaccines. Vaccine, 19(17-19), 2666-2672. http://dx.doi.org/10.1016/S0264-410X(00)00498-9. PMid:11257407.
Chauhan, G., Adams, H. H. H., Satizabal, C. L., Bis, J. C., Teumer, A., Sargurupremraj, M., Hofer, E., Trompet, S., Hilal, S., Smith, A. V., Jian, X., Malik, R., Traylor, M., Pulit, S. L., Amouyel, P., Mazoyer, B., Zhu, Y. C., Kaffashian, S., Schilling, S., Beecham, G. W., Montine, T. J., Schellenberg, G. D., Kjartansson, O., Guðnason, V., Knopman, D. S., Griswold, M. E., Windham, B. G., Gottesman, R. F., Mosley, T. H., Schmidt, R., Saba, Y., Schmidt, H., Takeuchi, F., Yamaguchi, S., Nabika, T., Kato, N., Rajan, K. B., Aggarwal, N. T., De Jager, P. L., Evans, D. A., Psaty, B. M., Rotter, J. I., Rice, K., Lopez, O. L., Liao, J., Chen, C., Cheng, C. Y., Wong, T. Y., Ikram, M. K., van der Lee, S. J., Amin, N., Chouraki, V., DeStefano, A. L., Aparicio, H. J., Romero, J. R., Maillard, P., DeCarli, C., Wardlaw, J. M., Hernández, M. D. C. V., Luciano, M., Liewald, D., Deary, I. J., Starr, J. M., Bastin, M. E., Muñoz Maniega, S., Slagboom, P. E., Beekman, M., Deelen, J., Uh, H. W., Lemmens, R., Brodaty, H., Wright, M. J., Ames, D., Boncoraglio, G. B., Hopewell, J. C., Beecham, A. H., Blanton, S. H., Wright, C. B., Sacco, R. L., Wen, W., Thalamuthu, A., Armstrong, N. J., Chong, E., Schofield, P. R., Kwok, J. B., van der Grond, J., Stott, D. J., Ford, I., Jukema, J. W., Vernooij, M. W., Hofman, A., Uitterlinden, A. G., van der Lugt, A., Wittfeld, K., Grabe, H. J., Hosten, N., von Sarnowski, B., Völker, U., Levi, C., Jimenez-Conde, J., Sharma, P., Sudlow, C. L. M., Rosand, J., Woo, D., Cole, J. W., Meschia, J. F., Slowik, A., Thijs, V., Lindgren, A., Melander, O., Grewal, R. P., Rundek, T., Rexrode, K., Rothwell, P. M., Arnett, D. K., Jern, C., Johnson, J. A., Benavente, O. R., Wasssertheil-Smoller, S., Lee, J. M., Wong, Q., Mitchell, B. D., Rich, S. S., McArdle, P. F., Geerlings, M. I., van der Graaf, Y., de Bakker, P. I. W., Asselbergs, F. W., Srikanth, V., Thomson, R., McWhirter, R., Moran, C., Callisaya, M., Phan, T., Rutten-Jacobs, L. C. A., Bevan, S., Tzourio, C., Mather, K. A., Sachdev, P. S., van Duijn, C. M., Worrall, B. B., Dichgans, M., Kittner, S. J., Markus, H. S., Ikram, M. A., Fornage, M., Launer, L. J., Seshadri, S., Longstreth Junior, W. T., & Debette, S., & Stroke Genetics Network (SiGN), the International Stroke Genetics Consortium (ISGC), METASTROKE, Alzheimer’s Disease Genetics Consortium (ADGC), and the Neurology Working Group of the Cohorts for Heart and Aging Research in Genomic Epidemiology (CHARGE) Consortium (2019). Genetic and lifestyle risk factors for MRI-defined brain infarcts in a population-based setting. Neurology, 92(5), 486-503. http://dx.doi.org/10.1212/WNL.0000000000006851. PMid:30651383.
Conceição, F. R., Moreira, Â. N., & Dellagostin, O. A. (2006). A recombinant chimera composed of R1 repeat region of Mycoplasma hyopneumoniae P97 adhesin with Escherichia coli heat-labile enterotoxin B subunit elicits immune response in mice. Vaccine, 24(29-30), 5734-5743. http://dx.doi.org/10.1016/j.vaccine.2006.04.036. PMid:16730864.
Conforti, V. A., de Avila, D. M., Cummings, N. S., Zanella, R., Wells, K. J., Ülker, H., & Reeves, J. J. (2008). CpG motif-based adjuvant as a replacement for Freund’s complete adjuvant in a recombinant LHRH vaccine. Vaccine, 26(7), 907-913. http://dx.doi.org/10.1016/j.vaccine.2007.12.010. PMid:18201806.
Dummer, L. A., Araujo, I. L., Finger, P. F., dos Santos Junior, A. G., da Rosa, M. C., Conceição, F. R., Fischer, G., van Drunen Littel-van den Hurk, S., & Leite, F. P. (2014). Immune responses of mice against recombinant bovine herpesvirus 5 glycoprotein D. Vaccine, 32(21), 2413-2419. http://dx.doi.org/10.1016/j.vaccine.2014.03.011. PMid:24657716.
Ferro, V. A., & Stimson, W. H. (1998). Investigation into suitable carrier molecules for use in an anti-gonadotrophin releasing hormone vaccine. Vaccine, 16(11-12), 1095-1102. http://dx.doi.org/10.1016/S0264-410X(98)80104-7. PMid:9682364.
Ferro, V., Khan, M. A., McAdam, D., Colston, A., Aughey, E., Mullen, A., Waterston, M., & Harvey, M. J. (2004). Efficacy of an antifertility vaccine based on mammalian gonadotrophin releasing hormone (GnRH-I): A histological comparison in male animals. Veterinary Immunology and Immunopathology, 101(1-2), 73-86. http://dx.doi.org/10.1016/j.vetimm.2004.03.011. PMid:15261694.
Gil, L. A., Cunha, C. E. P., Moreira, G. M. S. G., Salvarani, F. M., Assis, R. A., Lobato, F. C. F., Mendonça, M., Dellagostin, O. A., & Conceição, F. R. (2013). Production and evaluation of a recombinant chimeric vaccine against Clostridium botulinum Neurotoxin Types C and D. PLoS One, 8(7), 1-10. http://dx.doi.org/10.1371/journal.pone.0069692. PMid:23936080.
Han, X., Ren, X., Zeng, Y., Zhou, Y., Song, T., Cao, X., Du, X., Meng, F., Tan, Y., Liu, Y., Feng, J., Chu, M., & Zeng, X. (2016). Physiological interactions between the hypothalamic-pituitary-gonadal axis and spleen in rams actively immunized against GnRH. International Immunopharmacology, 38, 275-283. http://dx.doi.org/10.1016/j.intimp.2016.06.011. PMid:27322522.
Herbert, C. A., & Trigg, T. E. (2005). Applications of GnRH in the control and management of fertility in female animals. Animal Reproduction Science, 88(1-2), 141-153. http://dx.doi.org/10.1016/j.anireprosci.2005.05.007. PMid:16102921.
Higgins, D. (2004). Subcutaneous injection. Nursing Times, 100(50), 32-33. PMid:15633841.
Hsu, C. T., Ting, C. Y., Ting, C. J., Chen, T. Y., Lin, C. P., Whang-Peng, J., & Hwang, J. (2000). Vaccination against gonadotropin-releasing hormone (GnRH) using toxin receptor-binding domain-conjugated GnRH repeats. Cancer Research, 60(14), 3701-3705. PMid:10919636. Hunter, J. (2008). Intramuscular injection techniques. Nursing Standard, 22(24), 35-40. http://dx.doi.org/10.7748/ns2008.02.22.24.35. c6413. PMid:18318317.
Jegerlehner, A., Maurer, P., Bessa, J., Hinton, H. J., Kopf, M., & Bachmann, M. F. (2007). TLR9 signaling in B cells determines class switch recombination to IgG2a. Journal of Immunology (Baltimore, Md.: 1950), 178(4), 2415-2420. http://dx.doi.org/10.4049/jimmunol.178.4.2415. PMid:17277148.
Kaeberle, M. L. (1986). Function of carriers and adjuvants in induction of immune responses. In R. M. Nervig, P. M. Gough, M. L. Kaeberle & C. A. Whetstone (Eds.), Advances in Carriers and Adjuvants for Veterinary Biologics (pp. 11-23). Iowa State University Press.
Khan, M. A. H., Ferro, V. A., Koyama, S., Kinugasa, Y., Song, M., Ogita, K., Tsutsui, T., Murata, Y., & Kimura, T. (2007). Immunisation of male mice with a plasmid DNA vaccine encoding gonadotrophin releasing hormone (GnRH-I) and T-helper epitopes suppresses fertility in vivo. Vaccine, 25(18), 3544-3553. http://dx.doi.org/10.1016/j.vaccine.2007.01.089. PMid:17316929.
Nashar, T. O., Betteridge, Z. E., & Mitchell, R. N. (2001). Evidence for a role of ganglioside GM1 in antigen presentation: Binding enhances presentation of Escherichia coli enterotoxin B subunit (EtxB) to CD4(+) T cells. International Immunology, 13(4), 541-551. http://dx.doi.org/10.1093/intimm/13.4.541. PMid:11282993.
Orr, M. T., Fox, C. B., Baldwin, S. L., Sivananthan, S. J., Lucas, E., Lin, S., Phan, T., Moon, J. J., Vedvick, T. S., Reed, S. G., & Coler, R. N. (2013). Adjuvant formulation structure and composition are critical for the development of an effective vaccine against tuberculosis. Journal of Controlled Release, 172(1), 190-200. http://dx.doi.org/10.1016/j.jconrel.2013.07.030. PMid:23933525.
Pinheiro, A. F., Roloff, B. C., da Silveira Moreira, A., Berne, M. E. A., Silva, R. A., & Leite, F. P. L.. (2018). Identification of suitable adjuvant for vaccine formulation with the Neospora caninum antigen NcSRS2. Vaccine, 2018(36), 1154-1159. http://dx.doi.org/10.1016/j.vaccine.2018.01.051. PMid:29395519.
Ramos, C. R. R., Abreu, P. A. E., Nascimento, A. L. T. O., & Ho, P. L. (2004). A high-copy T7 Escherichia coli expression vector for the production of recombinant proteins with a minimal N-terminal His-tagged fusion peptide. Brazilian Journal Medical and Biological Research, 37(8), 1103-1109. https://doi.org/10.1590/S0100-879X2004000800001.
Richards, C. M., Aman, A. T., Hirst, T. R., Hill, T. J., & Williams, N. A. (2001). Protective mucosal immunity to ocular herpes simplex virus Type 1 infection in mice by using Escherichia coli Heat-Labile Enterotoxin B subunit as an adjuvant. Journal of Virology, 75(4), 1664-1671. http://dx.doi.org/10.1128/JVI.75.4.1664-1671.2001. PMid:11160664.
Sad, S., Gupta, H. M., Talwar, G. P., & Raghupathy, R. (1991). Carrierinduced suppression of the antibody response to a “self” hapten. Immunology, 74(2), 223-227. PMid:1748470.
Siel, D., Vidal, S., Sevilla, R., Paredes, R., Carvallo, F., Lapierre, L., Maino, M., Pérez, O., & Sáenz, L. (2016). Effectiveness of an immunocastration vaccine formulation to reduce the gonadal function in female and male mice by Th1/Th2 immune response. Theriogenology, 86(6), 1589-1598. http://dx.doi.org/10.1016/j.theriogenology.2016.05.019. PMid:27344434.
Smith, A. (2015). Subcutaneous injection. Nursing Standard, 29(38), 61. http://dx.doi.org/10.7748/ns.29.38.61.s43. PMid:25990183.
Tanghe, A., Denis, O., Lambrecht, B., Motte, V., van den Berg, T., & Huygen, K. (2000). Tuberculosis DNA vaccine encoding Ag85A is immunogenic and protective when administered by intramuscular needle injection but not by epidermal gene gun bombardment. Infection and Immunity, 68(7), 3854-3860. http://dx.doi.org/10.1128/IAI.68.7.3854-3860.2000. PMid:10858194.
Williams, N. A. (2000). Immune modulation by the cholera-like enterotoxin B-subunits: From adjuvant to immunotherapeutic. International Journal of Medical Microbiology, 290(4-5), 447-453. http://dx.doi.org/10.1016/S1438-4221(00)80062-4. PMid:11111925. Wilson-Welder, J. H., Torres, M. P., Kipper, M. J., Mallapragada, S. K., Wannemuehler, M. J., & Narasimhan, B. (2009). Vaccine adjuvants: Current challenges and future approaches. Journal of Pharmaceutical Sciences, 98(4), 1278-1316. http://dx.doi.org/10.1002/jps.21523. PMid:18704954.
Yamamoto, M., McGhee, J. R., Hagiwara, Y., Otake, S. & Kiyono, H. (2001). Genetically manipulated bacterial toxin as a new generation mucosal adjuvant. Scandinavian Journal of Immunology, 53(3), 211-217. http://dx.doi.org/10.1046/j.1365-3083.2001.00883.x.
Yamshchikov, G. V. Y., Barnd, D. L. B., Eastham, S. E., Galavotti, H. G., Patterson, J. W., Deacon, D. H. D., Teates, D., Neese, P., Grosh, W. W., Petroni, G., Engelhard, V. H., & Slingluff Junior, C. L. (2001). Evaluation of peptide vaccine immunogenicity in draining lymph nodes and peripheral blood of melanoma patients. International Journal of Cancer, 92(5), 703-711. http://dx.doi.org/10.1002/1097-0215(20010601)92:5<703::AID-IJC1250>3.0.CO;2-5. PMid:11340576.
Zhou, W. Y., Shi, Y., Wu, C., Zhang, W. J., Mao, X. H., Guo, G., Li, H. X., & Zou, Q. M. (2009). Therapeutic efficacy of a multi-epitope vaccine against Helicobacter pylori infection in BALB/c mice model. Vaccine, 27(36), 5013-5019. http://dx.doi.org/10.1016/j.vaccine.2009.05.009. PMid:19446591.
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