Miejscowa i ogólnoustrojowa odpowiedź immunologiczna na wybrane, rekombinowane antygeny bakteryjne (rHsp60 i rOMP40 Histophilus somni) u cieląt ras mlecznych

<p>In this study, we investigated early immunizations against selected conserved bacterial antigens. Calves were vaccinated twice intranasally and then subcutaneously with Histophilus somni recombinant proteins (rOMP40, rHsp60) mixed with one of two adjuvants: CpG ODN2007 or MPLA. The control group (Con) was injected with PBS. There were 11 calves in each group (experimental group I- CpG; experimental group II- MPLA; and control-Con). The first immunization was done between 24-48 hours of life and then twice in two weeks intervals. During experiment blood samples, nasal secretions and saliva were taken five times: S1- Immediately before the first intranasal immunization (24-48 hours of life). S2- Two weeks after the first immunization (14-16 days of age) and immediately before the second intranasal immunization. S3- Two weeks after the second immunization (28-30 days of age), immediately before the third subcutaneous immunization. S4- Two weeks after the third immunization (42-44 days of age). S5- two weeks after S4 (59-61 days of age). Before each sampling, clinical examination of the calves was performed (rectal temperature oC nasal discharge, cough, hearth, and breathing rates). In serum samples it was determined:  the reactivity of anti-rHsp60 and anti-rOMP40 antibodies in IgG1, IgG2, IgM and IgA classes  immunoglobulins (IgG1, IgG2, IgM and IgA classes) concentrations  acute phase proteins (haptoglobin, SAA, fibrinogen) concentrations In nasal secretions it was determined:  the reactivity of anti-rHsp60 and anti-rOMP40 antibodies in IgG1 and IgA classes  immunoglobulins (IgG1 and IgA classes) concentrations In whey colostrum it was determined:  the reactivity of anti-rHsp60 and anti-rOMP40 antibodies in IgG1, IgG2, IgM and IgA classes  immunoglobulins (IgG1, IgG2, IgM and IgA classes) concentrations In calf serum from S1, S3 and S5, the antibody against Bovine Herpesvirus-1 (BHV-1 ), Bovine Viral Diarrhea Virus (BVDV ), Bovine Respiratory Syncytial Virus (BRSV ), Bovine Parainfluenza-3 virus (PI-3V ), and Mycoplasma bovis were evaluated.</p>

本研究针对筛选得到的保守型细菌抗原开展了犊牛早期免疫接种研究。试验犊牛先接受两次经鼻内免疫，随后接受一次皮下免疫，疫苗为混合了两种佐剂之一的牛嗜眠嗜血杆菌（Histophilus somni）重组蛋白（rOMP40、rHsp60），两种佐剂分别为CpG ODN2007与单磷酰脂质A（MPLA）；对照组（Con组）则注射磷酸盐缓冲液（Phosphate Buffered Saline, PBS）。每组各设11头犊牛，分别为实验组I（佐剂为CpG ODN2007）、实验组II（佐剂为MPLA）以及对照组Con。
首次免疫于犊牛出生后24~48小时内完成，后续以两周为间隔开展后续免疫。试验期间共采集5次样本，各采样时间点如下：
S1：首次经鼻内免疫前（犊牛出生后24~48小时）；
S2：首次免疫后两周（日龄14~16天），即第二次经鼻内免疫前；
S3：第二次免疫后两周（日龄28~30天），即第三次皮下免疫前；
S4：第三次免疫后两周（日龄42~44天）；
S5：S4时间点后两周（日龄59~61天）。
每次采样前均对犊牛进行临床检查，检测指标包括直肠体温、鼻分泌物情况、咳嗽症状、心率及呼吸频率。
各样本的检测项目如下：
1. 血清样本：
✓ 抗rHsp60、抗rOMP40抗体在IgG1、IgG2、IgM、IgA类别的反应性
✓ 各类免疫球蛋白（IgG1、IgG2、IgM、IgA）的浓度
✓ 急性期蛋白（结合珠蛋白、血清淀粉样蛋白A（SAA）、纤维蛋白原）的浓度
2. 鼻分泌物样本：
✓ 抗rHsp60、抗rOMP40抗体在IgG1、IgA类别的反应性
✓ 各类免疫球蛋白（IgG1、IgA）的浓度
3. 初乳乳清样本：
✓ 抗rHsp60、抗rOMP40抗体在IgG1、IgG2、IgM、IgA类别的反应性
✓ 各类免疫球蛋白（IgG1、IgG2、IgM、IgA）的浓度
针对S1、S3、S5时间点采集的犊牛血清，额外评估了针对以下病原的抗体水平：牛疱疹病毒1型（Bovine Herpesvirus-1, BHV-1）、牛病毒性腹泻病毒（Bovine Viral Diarrhea Virus, BVDV）、牛呼吸道合胞体病毒（Bovine Respiratory Syncytial Virus, BRSV）、牛副流感3型病毒（Bovine Parainfluenza-3 Virus, PI-3V）以及牛支原体（Mycoplasma bovis）。