Construction, purification, and immunogenicity of recombinant cystein-cystein type chemokine receptor 5 vaccine.
Cystein-Cystein type chemokine receptor 5 (CCR5) is a seven-transmembrane, G-protein coupled receptor. It is a major coreceptor with CD4 glycoprotein mediating cellular entry of CCR5 strains of HIV-1. A lack of cell-surface expression of CCR5 found in the homozygous Delta32 CCR5 mutation, upregulation of CC chemokines and antibodies to CCR5 are associated with resistance to HIV infection. In addition, CCR5 can be blocked by three CC chemokines and antibodies to three extracellular domains of CCR5. Consequently, CCR5 is considered an attractive therapeutic target against HIV infection. In the current study, we constructed a recombinant vaccine by coupling a T helper epitope AKFVAAWTLKAA (PADRE) to the N terminus of CCR5 extracellular domains (PADRE-CCR5) and expressed this protein in Escherichia coli. We have developed an inexpensive and scalable purification process for the fusion protein from inclusion bodies and the final yields of 6mg purified fusion protein per gram of cell paste was obtained. The immunogenicity of the recombinant vaccine generated was examined in BALB/c mice. Sera from the vaccinated mice demonstrated high-titer specific antibodies to the recombinant vaccine, suggesting that PADRE-rCCR5 may be used as a candidate of active CCR5 vaccine.
Wu
Kongtian
K
Biotechnology Center of The Fourth Military Medical University, 17 Changle West Road, 710032 Xi'an, PR China.
Xue
Xiaochang
X
Wang
Zenglu
Z
Yan
Zhen
Z
Shi
Jihong
J
Han
Wei
W
Zhang
Yingqi
Y
D000818:Animals; D000906:Antibodies / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D019713:Receptors, CCR5 / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism*; D014611:Vaccination
D000906:Antibodies; D019713:Receptors, CCR5