IMMUNOHISTOLOGICAL DETECTION OF HUMAN CYTOTOXIC SUPPRESSOR T-CELLS USING ANTIBODIES TO A CD8 PEPTIDE SEQUENCE
IMMUNOHISTOLOGICAL DETECTION OF HUMAN CYTOTOXIC SUPPRESSOR T-CELLS USING ANTIBODIES TO A CD8 PEPTIDE SEQUENCE
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DOI：
10.1136/jcp.45.12.1084
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发表时间：
1992-12-01
影响因子：
3.4
通讯作者：
BROWN, MH
中科院分区：
文献类型：
--
作者：
MASON, DY;CORDELL, JL;BROWN, MH
目的：为了评估是否可以使用合成的CD 8肽序列的抗体在石蜡包埋的人组织样品中检测到细胞毒性/抑制性T细胞。方法：针对来自人CD 8分子α链的细胞质部分的13个氨基酸的肽序列，产生多克隆和单克隆抗体。结果：当用放射性标记的T细胞通过免疫沉淀进行测试时，这些抗体特异性地检测到CD 8多肽的天然形式，并且在低温恒温器切片中给出了细胞毒性/抑制性T细胞的预期染色模式。在兔子中培养的多克隆抗体也可用于与单克隆抗体联合双标记CD 8。石蜡包埋组织中也可见CD 8阳性细胞。如果组织已固定在布因固定液中，则无需对切片进行预先处理即可实现。当组织已暴露于传统的福尔马林固定，初步微波treatment was required.Conclusions：这些研究结果提供了进一步的证据表明，抗体对白细胞相关抗原，能够反应石蜡包埋的组织，可以通过免疫与合成肽序列。
Aims: To evaluate whether cytotoxic/suppressor T cells can be detected in paraffin wax embedded human tissue samples using antibodies to a synthetic CD8 peptide sequence.Methods: Polyclonal and monoclonal antibodies were raised against a 13 amino acid peptide sequence from the cytoplasmic portion of the alpha chain of the human CD8 molecule.Results: These antibodies specifically detected the native form of the CD8 polypeptide when tested by immunoprecipitation with radiolabelled T cells, and gave the expected staining pattern for cytotoxic/suppressor T cells in cryostat sections. Being raised in rabbits, the polyclonal antibodies were also useful for double labelling for CD8 in conjunction with monoclonal antibodies. CD8 positive cells could also be detected in paraffin wax embedded tissues. This was achieved without prior treatment of the sections if the tissue had been fixed in Bouin's fixative. When tissues had been exposed to conventional formalin fixation, preliminary microwave treatment was required.Conclusions: These findings provide further evidence that antibodies against leucocyte associated antigens, capable of reacting on paraffin wax embedded tissue, can be produced by immunisation with synthetic peptide sequences.