ارزیابی مقایسهای ایمنی‌زایی واکسن کونژوگه پنوموکوک در دو سویه موش آزمایشگاهی BALB/c و DBA/2: مطالعه پیش‌بالینی
محورهای موضوعی :
آسیب شناسی درمانگاهی دامپزشکی
رامین فرهودی
1
,
دلارام درود
2
,
گلشید javdani
3
,
محمد حسین هدایتی
4
1 - استادیار بخش کنترل کیفیت، مجتمع تولیدی تحقیقاتی انستیتو پاستور ایران، تهران، ایران.
2 - استادیار مجتمع تحقیقاتی انستیتو پاستور ایران، تهران، ایران.
3 - استادیار مجتمع تحقیقاتی انستیتو پاستور ایران، تهران، ایران.
4 - دانش‌آموخته دکترای تخصصی، بخش کنترل‌کیفیت مجتمع تولیدی تحقیقاتی انستیتو پاستور ایران، تهران، ایران.
تاریخ دریافت : 1400/11/13
تاریخ پذیرش : 1401/03/11
تاریخ انتشار : 1401/03/01
کلید واژه:
اسب,
واکسن,
فلوسایتومتری,
استرپتوکوک پنومونیه,
چکیده مقاله :
استرپتوکوک پنومونیه به راحتی از انسان به حیوانات منتقل شده و باعث پنومونی به‌خصوص در اسب می‌شود. واکسیناسیون گسترده انسان از انتقال این بیماری به میزبان اسب جلوگیری می‌کند. ایمنی‌زایی یک واکسن توسط روش‌های مختلف ارزیابی می‌شود. هدف این مطالعه، طراحی آزمونی جهت سنجش ایمنی‌زایی واکسن کونژوگه پنوموکوک به‌صورت بخشی از مطالعات پیش‌بالینی تولید واکسن است. پس از کشت استرپتوکوک پنومونیه سروتیپ 19F در محیط بلاد آگار، کلونی‌های به‌دست آمده با استفاده از رنگ فلورسنس نشاندار شدند. از طرفی سرم موش‌های BALB/c و DBA/2 که با 3 دز واکسن کونژگه پنوموکوک ایمن شده بودند، به منظور تعیین آنتی‌بادی‌هایی با خاصیت بیگانه‌خواری علیه استرپتوکوک پنومونیه جمع‌آوری شد. پس از مجاورت رقت‌های سرمی با باکتری نشاندار، توانایی بیگانه‌خواری باکتری‌ها توسط اپسونین‌های موجود در سرم (اپسونوفاگوسیتوز) با اضافه کردن ماکروفاژ موشی توسط فلوسایتومتری خوانش شد. در هر دو سویه موش با کاهش رقت سرمی درصد سلول هایی در سرم که باکتری را فاگوسیتوز کردند کاهش پیدا کرد. تیتر اپسونوفاگوسیتیک در موش BALB/c 128 و در موش DBA/2 64 گزارش گردید. از طرفی نتایج فلوسایتومتری با نتایج آزمون شمارش دستی کلونی‌های زنده تفاوت معنی‌داری داشت (001/0p≤، 89/0r=). طبق نتایج مطالعه حاضر مشخص شد سویه موش BALB/c میزبان بهتری به منظور انجام تعیین کارایی واکسن است. از طرفی استفاده از روش فلوسایتومتری مزایای بیشتری نسبت به روش سنجش دستی دارد. در نتیجه نتایج این مطالعه ما را یک قدم دیگر به تولید واکسن موثر نزدیک‌تر می‌کند.
چکیده انگلیسی:
Streptococcus pneumoniae is easily transmitted from humans to animals and causes pneumonia, especially in horses. Extensive human vaccination prevents the transmission of the disease to the horse host. The immunogenicity of a vaccine is evaluated by various methods. The aim of this study was to design a test to evaluate the immunogenicity of pneumococcal conjugate vaccine as part of pre-clinical studies of vaccine production. After culturing Streptococcus pneumoniae serotype 19F in blood agar medium, the obtained colonies were labeled using fluorescence dye. On the other hand, the serum of BALB/c and DBA/2 mice immunized with Pneumococcal Conjugate Vaccine was collected to determine antibodies with phagocytosis properties against Streptococcus pneumoniae. After proximity of serum dilutions with labeled bacteria, the ability of bacteria to phagocytosis by serum opsonins (opsonophagocytosis) was read by adding mouse macrophage cells by flow cytometry. In both strains, the percentage of cells in the serum that phagocytosed the bacterium decreased with decreasing serum dilution. Opsonophagocytic titers were reported 128 in BALB/c mice and 64 in DBA/2 mice. On the other hand, flow cytometry results were significantly different from the results of manual colony count test (r = 0.89, p≤0.001). According to the results of the present study, the BALB/c strain of mice was a better host to determine the efficacy of the vaccine. Also, using flow cytometry method has more advantages than manual assay method. As a result, the data of this study bring us one step closer to producing an effective vaccine.
منابع و مأخذ:
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Chanter, N. (1997). Streptococci and enterococci as animal pathogens. Journal of Applied Microbiology Symposium Supplement, 83(S1): 100S-109S.
Chiavolini, D., Pozzi, G. and Ricci, S. (2008). Animal models of Streptococcus pneumonia Disease. Clinical Microbiology Reviews, 21(4): 666-685.
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Feldman, C. and Anderson R. (2020). Recent advances in the epidemiology and prevention of Streptococcus pneumoniae infections. F1000 Research, 7(9): F1000.
Gestur, V., Ingileif, J., Steinn, J. and Helgi, V. (1994). Opsonization and antibodies to capsular and cell wall polysaccharide of Streptococcus pneumoniae. The Journal of Infectious Diseases, 170(3): 592-599.
Ginders, M., Leschnik, M., Künzel, F., Kampner, D., Mikula, C., Steindl, G., et al. (2017). Characterization of Streptococcus pneumoniae isolates from Austrian companion animals and horses. Acta Veterinaria Scandinavica, 59(1): 79.
Jia, P., Dai, S., Wu, T. and Yang, S. (2021). New approaches to anticipate the risk of reverse zoonosis, 36(7): 580-590.
Martinez, J.E., Clutterbuck, E.A., Li, H., Romero-Steiner, S. and Carlone, G.M. (2006). Evaluation of multiplex flow cytometric opsonophagocytic assays for determination of functional anticapsular antibodies to Streptococcus pneumoniae. Clinical and Vaccine Immunology, 13(4): 459-466.
Martinez, J.E., Romero-Steiner, S., Pilishvili, T., Barnard, S., Schinsky, J., Goldblatt, D., et al. (1999). A flowcytometric opsonophagocytic assay for measurement of functional antibodies elicited after vaccination with 23-valent pneumococcal polysaccharide vaccine. Clinical and Diagnostic Laboratory Immunology, 6(4): 581-586.
Masomian, M., Ahmad, Z., Gew, L.T. and Poh, C.L. (2020). Development of next generation Streptococcus pneumoniae vaccines conferring broad protection. Vaccines (Basel), 17; 8(1): 132.
Nau, R., Kaye, K., Sachdeva, M., Sande, E. and Tauber, M.G. (1994). Rifampin for therapy of experimental pneumococcal meningitis in rabbits. Antimicrobial Agents and Chemotherapy, 38: 1186-1189.
Papadatou, I., Tzovara, I. and Licciardi, P.V. (2019). The role of serotype-specific immunological memory in pneumococcal vaccination: Current knowledge and future prospects. Vaccines, 7(1): 13.
Park, C., Kwon, E.Y., Choi, S.M., Cho, S.Y., Byun, J.H., Park, J.Y., et al. (2017). Comparative evaluation of a newly developed 13-valent pneumococcal conjugate vaccine in a mouse model. Human Vaccines & Immunotherapeutics, 13(5): 1169-1176.
Robert, L.B. and Moon, H.N. (2006). Development and validation of a fourfold multiplexed opsonization assay (MOPA4) for pneumococcal antibodies. Clinical and Vaccine Immunology, 13(9): 1004-1009.
Romero-Steiner, S., Frasch, C., Concepcion, N., Goldblatt, D., Kayhty, H., Vakevainen, M., et al. (2003). Multilaboratory evaluation of a viability assay for measurement of opsonophagocytic antibodies specific to the capsular polysaccharides of Streptococcus pneumoniae. Clinical and Diagnostic Laboratory Immunology, 10(6): 415-422.
Saeland, E., Vidarsson, G. and Jonsdottir, I. (2000). Pneumococcal pneumonia and bacteremia model in mice for the analysis of protective antibodies. Microbial Pathogenesis, 29(2): 81-91.
Soheyli, Z., Soleimani, M. and Majidzadeh, K. (2017). A PCR Assay for detection of mycoplasma contamination in cell culture by rRNA 16S specific primers. Paramedical Sciences and Military Health, 12(2): 13-20. [In Persian]
Sorensen, R.U. and Edgar, D. (2019). Specific antibody deficiencies in clinical practice. The Journal of Allergy and Clinical Immunology: In Practice, 7(3): 801-808.
Stern, P.L. (2020). Key steps in vaccine development. Annals of Allergy, Asthma & Immunology, 125(1): 17-27.
Timoney, J.F. (2004). The pathogenic equine streptococci. Veterinary Research, 35(4): 397-409.
Turner, A.E.B., Gerson, J.E., So, H.Y., Krasznai, D.J., Hilaire, A.J.S. and Gerson, D.F. (2017). Novel polysaccharide-protein conjugates provide an immunogenic 13-valent pneumococcal conjugate vaccine for S. pneumonia. Synthetic and Systems Biotechnology, 2(1): 49-58.
Whatmore, A.M., King, S.J., Doherty, N.C., Sturgeon, D., Chanter, N. and Dowson, C.G. (1999). Molecular characterization of equine isolates of Streptococcus pneumoniae: natural disruption of genes encoding the virulence factors pneumolysin and autolysin. Infection and Immunity, 67(6): 2776-2782.
Woudenberg, I.A., Hoenderop, J.Y. and Michel, M.F. (1979). Efficacy of antimicrobial chemotherapy in experimental rat pneumonia: effects of impaired phagocytosis. Infection and Immunity Journal, 25(1): 366-375.
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Berical, A.C., Harris, D., Dela Cruz, C.S. and Possick, J.D. (2016). Pneumococcal Vaccination Strategies an Update and Perspective. Annals of American Thoracic Society, 13(6): 933-944.
Borsa, N., Pasquale, M.D. and Restrepo, M.I. (2019). Animal Models of pneumococcal pneumonia. International Journal of Molecular Science, 20(17): 4222.
Chanter, N. (1997). Streptococci and enterococci as animal pathogens. Journal of Applied Microbiology Symposium Supplement, 83(S1): 100S-109S.
Chiavolini, D., Pozzi, G. and Ricci, S. (2008). Animal models of Streptococcus pneumonia Disease. Clinical Microbiology Reviews, 21(4): 666-685.
Converso, T.R., Assoni, L., André, G.O., Darrieux, M. and Leite, L.C.C. (2020). The long search for a serotype independent pneumococcal vaccine. Expert Review of Vaccines, 19(1): 57-70.
Daniels, C.C., Rogers, P.D. and Shelton, C.M. (2016). Review of pneumococcal vaccines: Current polysaccharide vaccine recommendations and future protein antigenes. The Journal of Pediatric Pharmacology and Therapeutics, 21(1): 27-35.
Dockrell, H.D., Whyte Moira, M.K.B. and Timothy, J.M. (2012). Pneumococcal pneumonia mechanisms of infection and resolution. Chest, 142(2): 482-491.
Feldman, C. and Anderson R. (2020). Recent advances in the epidemiology and prevention of Streptococcus pneumoniae infections. F1000 Research, 7(9): F1000.
Gestur, V., Ingileif, J., Steinn, J. and Helgi, V. (1994). Opsonization and antibodies to capsular and cell wall polysaccharide of Streptococcus pneumoniae. The Journal of Infectious Diseases, 170(3): 592-599.
Ginders, M., Leschnik, M., Künzel, F., Kampner, D., Mikula, C., Steindl, G., et al. (2017). Characterization of Streptococcus pneumoniae isolates from Austrian companion animals and horses. Acta Veterinaria Scandinavica, 59(1): 79.
Jia, P., Dai, S., Wu, T. and Yang, S. (2021). New approaches to anticipate the risk of reverse zoonosis, 36(7): 580-590.
Martinez, J.E., Clutterbuck, E.A., Li, H., Romero-Steiner, S. and Carlone, G.M. (2006). Evaluation of multiplex flow cytometric opsonophagocytic assays for determination of functional anticapsular antibodies to Streptococcus pneumoniae. Clinical and Vaccine Immunology, 13(4): 459-466.
Martinez, J.E., Romero-Steiner, S., Pilishvili, T., Barnard, S., Schinsky, J., Goldblatt, D., et al. (1999). A flowcytometric opsonophagocytic assay for measurement of functional antibodies elicited after vaccination with 23-valent pneumococcal polysaccharide vaccine. Clinical and Diagnostic Laboratory Immunology, 6(4): 581-586.
Masomian, M., Ahmad, Z., Gew, L.T. and Poh, C.L. (2020). Development of next generation Streptococcus pneumoniae vaccines conferring broad protection. Vaccines (Basel), 17; 8(1): 132.
Nau, R., Kaye, K., Sachdeva, M., Sande, E. and Tauber, M.G. (1994). Rifampin for therapy of experimental pneumococcal meningitis in rabbits. Antimicrobial Agents and Chemotherapy, 38: 1186-1189.
Papadatou, I., Tzovara, I. and Licciardi, P.V. (2019). The role of serotype-specific immunological memory in pneumococcal vaccination: Current knowledge and future prospects. Vaccines, 7(1): 13.
Park, C., Kwon, E.Y., Choi, S.M., Cho, S.Y., Byun, J.H., Park, J.Y., et al. (2017). Comparative evaluation of a newly developed 13-valent pneumococcal conjugate vaccine in a mouse model. Human Vaccines & Immunotherapeutics, 13(5): 1169-1176.
Robert, L.B. and Moon, H.N. (2006). Development and validation of a fourfold multiplexed opsonization assay (MOPA4) for pneumococcal antibodies. Clinical and Vaccine Immunology, 13(9): 1004-1009.
Romero-Steiner, S., Frasch, C., Concepcion, N., Goldblatt, D., Kayhty, H., Vakevainen, M., et al. (2003). Multilaboratory evaluation of a viability assay for measurement of opsonophagocytic antibodies specific to the capsular polysaccharides of Streptococcus pneumoniae. Clinical and Diagnostic Laboratory Immunology, 10(6): 415-422.
Saeland, E., Vidarsson, G. and Jonsdottir, I. (2000). Pneumococcal pneumonia and bacteremia model in mice for the analysis of protective antibodies. Microbial Pathogenesis, 29(2): 81-91.
Soheyli, Z., Soleimani, M. and Majidzadeh, K. (2017). A PCR Assay for detection of mycoplasma contamination in cell culture by rRNA 16S specific primers. Paramedical Sciences and Military Health, 12(2): 13-20. [In Persian]
Sorensen, R.U. and Edgar, D. (2019). Specific antibody deficiencies in clinical practice. The Journal of Allergy and Clinical Immunology: In Practice, 7(3): 801-808.
Stern, P.L. (2020). Key steps in vaccine development. Annals of Allergy, Asthma & Immunology, 125(1): 17-27.
Timoney, J.F. (2004). The pathogenic equine streptococci. Veterinary Research, 35(4): 397-409.
Turner, A.E.B., Gerson, J.E., So, H.Y., Krasznai, D.J., Hilaire, A.J.S. and Gerson, D.F. (2017). Novel polysaccharide-protein conjugates provide an immunogenic 13-valent pneumococcal conjugate vaccine for S. pneumonia. Synthetic and Systems Biotechnology, 2(1): 49-58.
Whatmore, A.M., King, S.J., Doherty, N.C., Sturgeon, D., Chanter, N. and Dowson, C.G. (1999). Molecular characterization of equine isolates of Streptococcus pneumoniae: natural disruption of genes encoding the virulence factors pneumolysin and autolysin. Infection and Immunity, 67(6): 2776-2782.
Woudenberg, I.A., Hoenderop, J.Y. and Michel, M.F. (1979). Efficacy of antimicrobial chemotherapy in experimental rat pneumonia: effects of impaired phagocytosis. Infection and Immunity Journal, 25(1): 366-375.
