Prevention of bovine theileriosis by vaccine

疫苗预防牛泰勒虫病

• 批准号: 63480094
• 负责人: TAKAHASHI Kiyoshi
• 金额: $ 4.16万
• 依托单位: Rakuno Gakuen University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63480094/
• 关键词: Bovine; Theileria sergenti; Sporozoite vaccine; Merozoite vaccine; Anti-idiotype vaccine; Cellular immunity; スポロゾイト合成ワクチン; 牛のタイレリア原虫感染の予防; DNAプローブ; コバルト照射による弱毒化; メロゾイトのモノクローナル抗体; スポロゾイトのモノクローナル抗体

1. Recombinant vaccine against the sporozoite Purification of sprozoite antigen was carried out, but sufficient volume for experiment could not separate from tick-salivary gland. In a preliminary experiment, antiserum against extracted sporozoite antigen were prepared, and antigenicity of the sporozoite was compared with merozoite antigen by immunoblot analysis. Major proteins of the sporozoite antigen were M.W. of 74 and 78 kD, 32 and 42 kD in the merozoite It was showed that there were cross reaction in the sporozoite and the merozoite antigen. Therefor three calves injected monoclonal antibody (MAb) that recognized a polypeptide with M.W. 32 kD of the merozoite antigen were challenged by the sporozoites to demonstrate possibility of cross-protective immunity. As a result, appearance and peak of parasitemia were delayed to compare with the control calves.But we will continue to try producing sporozoite-specific recombinant vaccine, because the merozoite MAb (32 kD) does not inhibit i … More nfection of the sporozoite.2. Recombinant vaccine against the merozoite A MAb against M.W. 32 kD of the merozoite intensively inhibited infection of the merozoite. We decided to produce recombinant vaccine against the merozoite, and complementary DNA libraries have now been prepared from the merozoite MRNA. This type vaccine could be ineffective if antigen diversity such as Babesia bovis and T. parva occurred in T. sergenti. We therefor prepared the merozoite DNA probes to detect antigen diversity of T. sergenti.3. Anti-idiotype vaccine Twenty two clones of monoclonal idiotype antibody against merozoite MAb paratope were prepared by routine methods. Three clones of them showed internal image by IFA.4. Cellular immune responses The macrophage activity temporarily increased prior to reach peak of parasitemia, and increased proliferation of T-cell coincided with peak of parasitemia following them NK activity increased. It was considered that activated macrophages by lymphokines acted as a main inhibitor of the parasite Less

1.对虫体抗原的子孢子进行了重组疫苗的纯化，但实验所需的体积不足以从唾液腺中分离出来。在初步实验中，制备了针对提取的子孢子抗原的抗血清，并通过免疫印迹分析对子孢子和裂殖子抗原的抗原性进行了比较。子孢子抗原的主要蛋白为74和78kD，32kD和42kD，说明子孢子和裂殖子抗原之间存在交叉反应。因此，用识别裂殖子抗原分子量为32kD的多肽的单抗(MAb)攻击3头小牛子孢子，以证明交叉保护免疫的可能性。因此，与对照牛相比，寄生虫血症的出现和高峰都延迟了。但我们将继续尝试生产针对子孢子特异性的重组疫苗，因为裂殖子单抗(32kD)不抑制I…子孢子感染较多。针对裂殖子32kD M.W.32kD的裂殖子A单抗重组疫苗能强烈抑制裂殖子的感染。我们决定生产针对裂殖子的重组疫苗，现在已经从裂殖子mRNA中制备了互补的DNA文库。如果瑟氏毛滴虫出现抗原多样性，如牛巴贝斯虫和微小毛滴虫，则这种类型的疫苗可能无效。为此，我们制备了裂殖子DNA探针，用于检测弓形虫的抗原多样性。抗独特型疫苗用常规方法制备22株抗裂殖子单抗独特型单抗。其中3个克隆经IFA检测显示有内部图像。细胞免疫反应：巨噬细胞活性在寄生虫血症高峰期前短暂升高，T细胞增殖增强与寄生虫血症高峰期一致，随后NK活性升高。目前认为淋巴因子激活的巨噬细胞是寄生虫的主要抑制因子。

项目成果

S. Shirakata et al: "Localization of surface antigen on Theileria sergenti merozoite by monoclonal antibodies." Jpn. J. Vet. Sci. 51, 831-833, 1989.

S. Shirakata 等人：“通过单克隆抗体对塞氏泰勒虫裂殖子上的表面抗原进行定位。”

N.KAJIWARA: "Specifie DNA probe for the diagnosis of Theileria sergenti infeetion in cattle" Jpn.J.Vet.Sci.(1990)

N.KAJIWARA：“用于诊断牛塞氏泰勒虫感染的特异性 DNA 探针”Jpn.J.Vet.Sci.(1990)

Y.Yasutomi et al: "Theileria sergenti infection induces proliferation of bovine lymphocytes" Jpn.J.Vet.Sci.(submitted). (1990)

Y.Yasutomi 等：“Theileria sergenti 感染诱导牛淋巴细胞增殖”Jpn.J.Vet.Sci.（已提交）。

S.SHIRAKATA: "Localization of serface antigen on Theileria sergenti merozoite by monoclonal antibodies" Jpn.J.Vet.Sci.51. 831-833 (1989)

S.SHIRAKATA：“通过单克隆抗体对瑟氏泰勒虫裂殖子上的 Serface 抗原进行定位”Jpn.J.Vet.Sci.51。

N.Sasaki et al: "Maturation of Theileria sergenti in salivary glands of Haemaphysalis longicornis induced by temperature stimulation" Jpn.J.Vet.Sci.52. inpress (1990)

N.Sasaki 等人：“温度刺激诱导长角血蜱唾液腺中瑟氏泰勒虫的成熟”Jpn.J.Vet.Sci.52。