Post-immunization immunohistochemical expression of Caspase 3 and p53 apoptotic markers in experimental hydatidosis

Abstract

The aim of this study was to investigate post-immunization apoptotic changes in experimental hydatidosis, using Caspase 3 and p53 immunohistochemical markers. Two groups of rabbits were immunized with a crude antigen (group 1) or a partially purified antigen (group 2) and were compared to an infected non-immunized control group. More effective immune responses were obtained in group 2 than group 1, signified by fewer and smaller cystic lesions and more severe destructive changes. Normal growth of cysts was attained in the control group, with no expression of apoptotic markers. Significantly higher expression of Caspase 3 and p53 were observed in group 1 compared to group 2, as indicated by OD and area percentage, respectively (Group 1 Caspase 3: 0.89±0.21, 93.5%±6.2; Group 1 p53: 0.46±0.18, 53.26%±11.6; Group 2 Caspase 3: 0.52±0.15, 49.23%±11.7; Group 2 p53: 0.19±0.4, 18.17%±7.3). Vaccine-induced immune responses and cellular damage may underlie the expression of apoptotic markers that appeared to result in a degenerative and atrophic course of action upon immunization. The results of the current study emphasize the importance of immunization for the stimulation of protective immune responses and in preventing mechanisms of evasion to ensure normal cell growth. A cost/benefit control program that implements proper vaccine preparations should be further assessed for complete elimination of severe infections in endemic areas.Keywords: Experimental hydatidosis, Caspase 3, p53, apoptosis. ResumoO objetivo do presente estudo foi investigar mudanças de apoptose pós-imunização em hidatidose experimental, usando os marcadores imuno-histoquímicos Caspase 3 e p53. Dois grupos de coelhos foram imunizados com antígeno bruto (grupo 1) e com antígeno parcialmente purificado (grupo 2). Estes grupos foram comparados a um grupo controle infectado e não-imunizado. Respostas imunes mais eficientes foram obtidas do grupo 2, que apresentou lesões císticas menores e menos frequentes, e mudanças destrutivas mais graves. Cistos cresceram normalmente no grupo controle, sem expressão dos marcadores de apoptose. Expressões significativamente mais altas de Caspase 3 e p53 foram observadas no grupo 1 quando comparado ao grupo 2, como indicado por DO e área de percentagem, respectivamente (Grupo 1 Caspase 3: 0,89±0,21, 93,5%±6,2; Grupo 1 p53: 0, 46±0,18, 53,26%±11,6; Grupo 2 Caspase 3: 0,52±0,15, 49,23%±11,7; Grupo 2 p53: 0,19±0,4, 18,17%±7,3). Respostas imunológicas induzidas por vacinas e danos celulares podem ser a base para a expressão dos marcadores de apoptose cujos desfechos demonstraram ação degenerativa e atrófica durante imunização. Os resultados do presente estudo enfatizam a importância da imunização para o estímulo de respostas imunes de proteção e para mecanismos de prevenção de evasão para garantir crescimento celular normal. Um programa de controle de custo/benefício que implemente preparações de vacinas adequadas deve ser analisado em mais detalhe para a completa eliminação de infecções graves em áreas endêmicas.Pal...

Interactions between hydatid cyst and regulated cell death may provide new therapeutic opportunities

“…Understanding the importance of these activations is critical because RCD is an integral part of health and disease and is stimulated by a variety of pathologic and physiologic stimuli. Histologic studies have shown that many factors are important for stimulating RCD in parasites and infected host cells [14,35,36]. A large number of studies have shown that the expression level of apoptosis-inducing factors in the germinal layer of sterile cysts is higher than that of fertile cysts, and the expression rate of anti-apoptotic factors in fertile cysts is higher, which may affect cystic infertility.…”

“…Based on that assumption, one could speculate that anti-cancer therapeutic approaches, including radiotherapy, should be tested to induce regulated cell death (RCD) of the parasite [25]. The second option is to address the issue of pro-apoptotic effects of HCF produced during infection of the intermediate host, and to test the hypothesis of potential value of certain specific compounds of this fluid as therapeutic agents for human cancers, including melanoma and breast cancer [11,14,16,39,51]. These two approaches may open an extensive research area, provided preliminary results obtained to date are consolidated and confirmed in large controlled studies.…”

A new experimental model of intrinsic denervation in ileum from wistar rats through intramural microinjections of benzalkonium chloride

“…The positive assessment of H2AX detects oxidative stress, which may result in breaking the double stranded DNA and thus begin the apoptosis process, evaluated by immunolabeling Caspase-3 (Berra et al 2006, Terzian et al 2007, Cleaver 2011, Redon et al 2012, Ghosal & Chen 2013, El-Aal et al 2016. For verification of the effectiveness of denervation by CB, attesting the neuronal death and determination of the period in which this event occurred, was evaluated in this study, the presence of severe damage to genetic material, by means of immunohistochemical technique for detection of cells positively marked to H2AX, effective scorer in the presence of the double-stranded breaks of DNA (Sak & Stuschke 2010, Cleaver 2011, Ghosal & Chen 2013, characterizing the oxidative stress and the presence of neuronal apoptosis through immunolabeling of Caspase-3 in different times (24, 48 hours, 30 and 90 days).…”

“…The results obtained in this study demonstrated the feasibility and effectiveness of intramural application of CB to get the digestive tube denervation, with a view to greater precision of this method of application of CB, through intramural applications. It was also possible to deepen the knowledge about the evolution of neuronal injury caused in this process, through histopathological analysis and immunohistochemistry by immunolabeling histones specific gamma-H2AX (H2AX) that assesses the presence of oxidative nuclear stress injuries (Berra et al 2006, Redon et al 2012) and Caspase-3, which detects apoptose (Terzian et al 2007, El-Aal et al 2016) of the myenterics neurons.…”

The Investigation of the Effect and Mechanism of Sophora moorcroftiana Alkaloids in Combination with Albendazole on Echinococcosis in an Experimental Rats Model

“…Echinococcus granulosus protoscolices were kindly provided by Qinghai Institute for Endemic Disease Prevention and Control, China. The protoscolices were aseptically removed from liver hydatid cysts obtained from cattle and washed several times with saline containing 1500 U/mL penicillin and 1000 U/mL streptomycin [ 11 ]. The survival rate of the protoscolices exceeded 95% after these procedures.…”