Neutralization Characteristics Of CRF01_AE Pseudovirus Of HIV-1 In China

HIV was identified as the pathogen of human immunodeficiency disease in 1983.More than 30 years after the first case of HIV infection was found in Africa,it has spread to all parts of the world and caused a pandemic.It has caused great impact on the world economy,health and social stability,and has attracted great attention from the World Health Organization and governments of various countries.Due to the high mutagenicity and recombination of HIV-1,different epidemic strains were produced and regional differences were gradually formed.CRF01_AE recombination derived from subtype A has become the most prevalent subtype in China.The lack of natural clearance of HIV-1 infection and the lack of clear immunological indicators related to HIV-1 protection have made the development of an HIV-1 vaccine diffic ? lt.Currently,there is no prophylactic HIV-1 vaccine that can induce sufficient protective effect.HIV-1 membrane protein envelope(ENV)plays a binding role with cell receptors in infection and contains epitopes recognized by neutralizing antibodies,which is the main target of neutralizing antibodies.The binding of neutralizing antibodies to ENV is very important for preventing HIV-1 infection.Pseudoviruses can replace live viruses in BSL-2 laboratory research experiments,has been widely used in virus research.Env protein can be inserted into the virus skeleton to form a functional pseudovirus with single round infection,which has been widely used in the evaluation of vaccines,antibodies and dr?gs.Pseudoviruses including B/B 'subtype,CRF07/08_BC subtype and some CRF01_AE subtype have been isolated and constructed from the plasma of infected patients in our laboratory.The epidemic trend of CRF01_AE is becoming more and more serious,and more and more mutant strains have emerged.Recently,our laboratory has constructed a library of 37 pseudoviruses of CRF01_AE subtype and 9 pseudoviruses of other minor epidemic subtypes.The new CRF01_AE subtype virus sequences were analyzed and found to be quite different from the existing sequences in the library,which co ? ld realize the expansion of the library.In order to further analyze the response characteristics of the virus to the Broader Neutralizing Monoclonal antibodies(bnmAbs),we studied the neutralization characteristics of 46 pseudoviruses against 11 bnmAbs.We also analyzed the epitopes recognized by these antibodies.The res?lts showed that the broad-spectrum monoclonal antibody recognizing CD4 binding sites(cd4bs)had the strongest neutralization ability to Pseudoviruses.VRC01 co?ld neutralize 100%(46 /46)of Pseudoviruses,and 80.4%(37 / 46)of Pseudoviruses co?ld be neutralized by3bnc117;The res ? lts showed that the Mc Ab recognizing the near membrane extracell ? lar domain(mper)co ? ld neutralize 67.4%(31 / 46)Pseudoviruses in4E10 and 76%(35 / 46)Pseudoviruses in 10e8;Detection of novel CRF01 by broad-spectrum monoclonal antibody recognizing V3 loop region_Pgt135 co?ld not even neutralize one strain of pseudovirus,and 10-1074 co?ld only neutralize 17.3%(8 / 46).Compared with the neutralization characteristics of the previously constructed B / B 'subtype pseudovirus,seven of the eight broad-spectrum monoclonal antibodies(VRC01,vrc03,Ig G1b12,2G12,2F5,4E10 and 10e8)were used for CRF01_AE Pseudoviruses are more diffic?lt to neutralize except PG9,The neutralization effect of the new sieve m Abs(such as VRC01 and 10E8)against pseudoviruses was also generally stronger than that of the old sieve m Abs(2F5 and Ig G1b12).In order to improve the detection level of vaccine neutralization antibody in China,the neutralization sensitivity of pseudoviruses to plasma disk was also divided into three categories according to the previous method,which were high,medium and low,and used to evaluate the level of humoral immune response caused by vaccine.In this study,by analyzing the neutralization sensitivity of pseudovirus CRF01_AE subtype to Bnmabs,we can s?ggest some effective treatment options for infected patients.By analyzing the recognition epitopes of Bnmabs,we can also provide information for the design of immunogens and vaccine evaluation of subtype CRF01_AE.The abundance of pseudovirus libraries and the calibration of newly constructed pseudoviruses also provide a relatively perfect platform for the evaluation of vaccines,therapeutic monoclonal antibodies and dr ? gs in the later stage,and provide support for the development and evaluation of vaccines in China.The newly constructed pseudovirus in this study has been used in the efficacy evaluation of some HIV-1 prophylactic vaccines.