Satoh Ryoichi, Kaku Ayumi, Satomura Megumi, Kohori Michiyo, Noura Kanako, Furukawa Tomoko, Kotake Masako, Takano Tomomi, Hohdatsu Tsutomu
Laboratory of Veterinary Infectious Disease, School of Veterinary Medicine, Kitasato University, Towada, Aomori 034-8628, Japan.
J Feline Med Surg. 2011 Jun;13(6):427-35. doi: 10.1016/j.jfms.2011.01.008. Epub 2011 Feb 22.
Feline infectious peritonitis virus (FIPV) can cause a lethal disease in cats, feline infectious peritonitis (FIP). The antibody-dependent enhancement (ADE) of FIPV infection has been recognised in experimentally infected cats, and cellular immunity is considered to play an important role in preventing the onset of FIP. To evaluate the importance of cellular immunity for FIPV infection, monoclonal antibodies (MAbs) against feline interferon (fIFN)-γ were first created to establish fIFN-γ detection systems using the MAbs. Six anti-fIFN-γ MAbs were created. Then, the difference in epitope which those MAbs recognise was demonstrated by competitive enzyme-linked immunosorbent assay (ELISA) and IFN-γ neutralisation tests. Detection systems for fIFN-γ (sandwich ELISA, ELISpot assay, and two-colour flow cytometry) were established using anti-fIFN-γ MAbs that recognise different epitopes. In all tests, fIFN-γ production from peripheral blood mononuclear cells (PBMCs) obtained from cats experimentally infected with an FIPV isolate that did not develop the disease was significantly increased by heat-inactivated FIPV stimulation in comparison with medium alone. Especially, CD8(+)fIFN-γ(+) cells, but not CD4(+)fIFN-γ(+) cells, were increased. In contrast, fIFN-γ production from PBMCs isolated from cats that had developed FIP and specific pathogen-free (SPF) cats was not increased by heat-inactivated FIPV stimulation. These results suggest that cellular immunity plays an important role in preventing the development of FIP. Measurement of fIFN-γ production with the anti-fIFN-γ MAbs created in this study appeared to be useful in evaluating cellular immunity in cats.
猫传染性腹膜炎病毒（FIPV）可在猫中引发一种致命疾病，即猫传染性腹膜炎（FIP）。FIPV感染的抗体依赖性增强（ADE）已在实验感染的猫中得到确认，并且细胞免疫被认为在预防FIP发病中起重要作用。为了评估细胞免疫对FIPV感染的重要性，首先制备了抗猫干扰素（fIFN）-γ单克隆抗体（MAb），以使用这些MAb建立fIFN-γ检测系统。制备了六种抗fIFN-γ MAb。然后，通过竞争性酶联免疫吸附测定（ELISA）和IFN-γ中和试验证明了这些MAb识别的表位差异。使用识别不同表位的抗fIFN-γ MAb建立了fIFN-γ检测系统（夹心ELISA、ELISpot测定和双色流式细胞术）。在所有试验中，与单独的培养基相比，用热灭活的FIPV刺激来自实验感染未发病的FIPV分离株的猫的外周血单个核细胞（PBMC），fIFN-γ的产生显著增加。特别是，CD8(+)fIFN-γ(+)细胞增加，而CD4(+)fIFN-γ(+)细胞未增加。相反，热灭活的FIPV刺激并未增加从患有FIP的猫和无特定病原体（SPF）猫分离的PBMC中fIFN-γ的产生。这些结果表明细胞免疫在预防FIP发展中起重要作用。用本研究中制备的抗fIFN-γ MAb测量fIFN-γ的产生似乎有助于评估猫的细胞免疫。
