Abstract:16/19) of the ALV-J layer isolates displayed less than 92.5% sequence homology to those of the ALV-J broiler isolates, although the transcriptional regulatory elements that are typical of avian retroviruses were highly conserved. Several unique nucleotide substitutions in the env gene, the U3 region, and the E element of most of the ALV-J layer isolates were detected. These results suggested that the env gene, E element, and U3 region in the ALV-J layer isolates have evolved rapidly and were significantly diff… Show more
“…Immunofluorescence assays showed that the mAb 4A3 could react with the British prototype strain HPRS-103 [1], the American strains ADOLHc1 and ADOL-7501 [27], the early Chinese broiler strain NX0101 [3], and recent Chinese layer strains (HuB09JY03, JL08CH3-1, SD09DP04, HLJ09SH01, LN08SY10) [21] (Fig. 2C).…”
Section: Production and Characterization Of Mabsmentioning
confidence: 94%
“…Because ALV-J has become widespread and has caused great economic losses to poultry in China [21], an effective and rapid diagnostic method for ALV-J infections is urgently needed. Defined B-cell epitopes, which are regions recognized by specific antibodies or B-cell receptors, have become useful tools for the development of diagnostic methods [22].…”
Avian leukosis virus subgroup J (ALV-J) is an avian oncogenic retrovirus that has caused severe economic losses in China. Gp85 protein is the main envelope protein and the most variable structural protein of ALV-J. It is also involved in virus neutralization. In this study, a specific monoclonal antibody, 4A3, was produced against the ALV-J gp85 protein. Immunofluorescence assays showed that 4A3 could react with different strains of ALV-J, including the British prototype isolate HPRS103, the American strains, an early Chinese broiler isolate, and layer isolates. A linear epitope on the gp85 protein was identified using a series of partially overlapping fragments spanning the gp85-encoding gene and subjecting them to western blot analysis. The results indicated that (134)AEAELRDFI(142) was the minimal linear epitope that could be recognized by mAb 4A3. Enzyme-linked immunosorbent assay (ELISA) revealed that chicken anti-ALV-J sera and mouse anti-ALV-J gp85 sera could also recognize the minimal linear epitope. Alignment analysis of amino acid sequences indicated that the epitope was highly conserved among 34 ALV-J strains. Furthermore, the epitope was not conserved among subgroup A and B of avian leukosis virus (ALV). Taken together, the mAb and the identified epitope may provide valuable tools for the development of new diagnostic methods for ALV-J.
“…Immunofluorescence assays showed that the mAb 4A3 could react with the British prototype strain HPRS-103 [1], the American strains ADOLHc1 and ADOL-7501 [27], the early Chinese broiler strain NX0101 [3], and recent Chinese layer strains (HuB09JY03, JL08CH3-1, SD09DP04, HLJ09SH01, LN08SY10) [21] (Fig. 2C).…”
Section: Production and Characterization Of Mabsmentioning
confidence: 94%
“…Because ALV-J has become widespread and has caused great economic losses to poultry in China [21], an effective and rapid diagnostic method for ALV-J infections is urgently needed. Defined B-cell epitopes, which are regions recognized by specific antibodies or B-cell receptors, have become useful tools for the development of diagnostic methods [22].…”
Avian leukosis virus subgroup J (ALV-J) is an avian oncogenic retrovirus that has caused severe economic losses in China. Gp85 protein is the main envelope protein and the most variable structural protein of ALV-J. It is also involved in virus neutralization. In this study, a specific monoclonal antibody, 4A3, was produced against the ALV-J gp85 protein. Immunofluorescence assays showed that 4A3 could react with different strains of ALV-J, including the British prototype isolate HPRS103, the American strains, an early Chinese broiler isolate, and layer isolates. A linear epitope on the gp85 protein was identified using a series of partially overlapping fragments spanning the gp85-encoding gene and subjecting them to western blot analysis. The results indicated that (134)AEAELRDFI(142) was the minimal linear epitope that could be recognized by mAb 4A3. Enzyme-linked immunosorbent assay (ELISA) revealed that chicken anti-ALV-J sera and mouse anti-ALV-J gp85 sera could also recognize the minimal linear epitope. Alignment analysis of amino acid sequences indicated that the epitope was highly conserved among 34 ALV-J strains. Furthermore, the epitope was not conserved among subgroup A and B of avian leukosis virus (ALV). Taken together, the mAb and the identified epitope may provide valuable tools for the development of new diagnostic methods for ALV-J.
“…The detection limit of this method was as low as 1 ϫ 10 2 viral DNA copies for each of the three plasmids. The mPCR method was compared with the virus isolation method because virus isolation is considered to be the gold standard for viral detection (10). In animal experiments, the mPCR detected ALVs 2 to 4 days earlier than virus isolation from whole-blood samples and 2 days earlier than virus isolation from cloaca swabs.…”
Section: Discussionmentioning
confidence: 99%
“…No field cases of ALV-J infection or tumors in layer chickens were observed worldwide until 2004 (9-11). However, parent and commercial layer flocks in China have experienced outbreaks of this virus in recent years, causing serious economic losses (10). Thus, ALV-A, ALV-B, and ALV-J are not only the most common but also the most dangerous viruses to the poultry industry.…”
“…As the spread of the disease is associated with diverse pathotypes and could result in enormous economic losses, ALV-J-induced disease in layer-type chickens has become one of the most important problems facing the global poultry industry. In addition to the fact that the higher incidence of lymphoid leukosis (LL) or erythroblastosis (EB) in susceptible chickens is induced by the ALV subgroups A (ALV-A) and B (ALV-B) (5), ALV-J was also found to be the main cause of ML and hemangioma after a long latent period, as well as a wide variety of other tumors at a lower incidence (6)(7)(8)(9). Further studies have found that virus-and/or cell lineage-specific determinants are also important for tumor development, but the oncogenicity of ALV-J remains unclear.…”
The integration of retroviruses into the host genome following nonrandom genome-wide patterns may lead to the deregulation of gene expression and oncogene activation near the integration sites. Slow-transforming retroviruses have been widely used to perform genetic screens for the identification of genes involved in cancer. To investigate the involvement of avian leukosis virus subgroup J (ALV-J) integration in myeloid leukosis (ML) in chickens, we utilized an ALV-J insertional identification platform based on hybrid capture target enrichment and next-generation sequencing (NGS). Using high-definition mapping of the viral integration sites in the chicken genome, 241 unique insertion sites were obtained from six different ALV-J-induced ML samples. On the basis of previous statistical definitions, MYC, TERT, and ZIC1 genes were identified as common insertion sites (CIS) of provirus integration in tumor cells; these three genes have previously been shown to be involved in the malignant transformation of different human cell types. Compared to control samples, the expression levels of all three CIS genes were significantly upregulated in chicken ML samples. Furthermore, they were frequently, but not in all field ML cases, deregulated at the mRNA level as a result of ALV-J infection. Our findings contribute to the understanding of the relationship between multipathotypes associated with ALV-J infection and the molecular background of tumorigenesis.IMPORTANCE ALV-Js have been successfully eradicated from chicken breeding flocks in the poultry industries of developed countries, and the control and eradication of ALV-J in China are now progressing steadily. To further study the pathogenesis of ALV-J infections, it will be necessary to elucidate the in vivo viral integration and tumorigenesis mechanism. In this study, 241 unique insertion sites were obtained from six different ALV-J-induced ML samples. In addition, MYC, TERT, and ZIC1 genes were identified as the CIS of ALV-J in tumor cells, which might be a putative "driver" for the activation of the oncogene. In addition, the CIS genes showed deregulated expression compared to nontumor samples. These results have potentially important implications for the mechanism of viral carcinogenesis.
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