Intramammary expression and therapeutic effect of a human lysozyme-expressing vector for treating bovine mastitis |
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Authors: | Huai-chang Sun Fang-ming Xue Ke Qian Hao-xia Fang Hua-lei Qiu Xin-yu Zhang Zhao-hua Yin |
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Institution: | (1) School of Veterinary Medicine, Yangzhou University, Yangzhou, 225009, China |
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Abstract: | To develop a gene therapy strategy for treating bovine mastitis, a new mammary-specific vector containing human lysozyme (hLYZ)
cDNA and kanamycin resistance gene was constructed for intramammary expression and clinical studies. After one time acupuncture
or intracisternal infusion of healthy cows with 400 μg of the p215C3LYZ vector, over 2.0 μg/ml of rhLYZ could be detected
by enzymatic assay for about 3 weeks in the milk samples. Western blotting showed that rhLYZ secreted into milk samples from
the vector-injected cows had molecular weight similar to that of the natural hLYZ in human colostrums. Twenty days after the
primary injection, the quarters were re-injected with the same vector by quarter acupuncture and even higher concentrations
of rhLYZ could be detected. Indirect competitive ELISA of milk samples showed that the vector injection did not induce detectable
humoral immune response against hLYZ. Clinical studies showed that twice acupuncture of quarters with the p215C3LYZ vector
had overt therapeutic effect on clinical and subclinical mastitis previously treated with antibiotics, including disappearance
of clinical symptoms and relatively high microbiological cure rates. These data provide a solid rationale for using the vector
to develop gene therapy for treating bovine mastitis.
Project (Nos. BJ2001315 and BE2004611) supported by the Department of Science and Technology of Jiangsu Province, China |
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Keywords: | Human lysozyme cDNA Mammary-specific vector Expression Mastitis Gene therapy |
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