Differential cellular immune responses to wild‐type and attenuated edmonston tag measles virus strains are primarily defined by the viral phosphoprotein gene

Abstract

The measles virus phosphoprotein (P) gene encodes the P, V, and C proteins, which have multiple functions including type I interferon (IFN) inhibition. With a focus on viral immune modulation, we conducted a study on healthy vaccinees (n = 179) to compare cytokine secretion patterns/cell frequencies and gene expression after in vitro encounter with a highly attenuated strain of measles virus (MVEdmtag), wild-type MV (MVwt) or recombinant MVEdmtag expressing the wild-type P gene (MVwtP). Cytokines were quantified by ELISA and Elispot. Gene expression profiling was performed using real-time PCR. We found differential MV-specific cytokine responses to all detected cytokines characterized by significantly higher cytokine levels (P <0.001) and higher frequencies (P <0.0001) of cytokine-producing cells after stimulation with the highly attenuated MVEdm-tag strain in comparison with MVwt or MVwtP. Furthermore, gene expression profiling revealed significant cytokine suppression at the transcriptional level for viruses encoding the functional wt P gene, compared to attenuated MVEdmtag (P <0.05). Using lentivirus-mediated stable expression of P gene-encoded proteins in human cell lines, we demonstrated that the expression of the functional wt V protein significantly down-modulated the induction of IFNs type I, II, and III in lymphocytes and monocytes. Taken together our results indicate that Th1, Th2, and innate/inflammatory cytokine responses in vaccinees are suppressed both at the protein and transcriptional level by viruses expressing the functional wt P gene products. The functional P gene-encoded viral proteins (particularly V proteins) emerge as crucial immune evasion factors for modulating and shaping the measles virus-specific cytokine responses in humans.

Associations between single nucleotide polymorphisms and haplotypes in cytokine and cytokine receptor genes and immunity to measles vaccination

“…Cytokine and cytokine receptors are intrinsically linked to the generation and regulation of adaptive immune responses, and therefore are reasonable targets for candidate gene-based genetic studies [6,29-32]. We have previously sampled a smaller number of candidate cytokine and cytokine receptor gene polymorphisms/SNPs (n=58) in Caucasians (n=118) and African-Americans (n=85) and reported initial moderate associations between SNPs in IL2, IL10, IL12B, IL4RA, IL12RB, IL6 and TNFA genes and measles vaccine-induced immunity [3,5].…”

Specific Elimination of CD133+ Tumor Cells with Targeted Oncolytic Measles Virus

“…Our data confirm that receptor attachment is not the crucial step for vaccine-derived measles viruses to infect HSC. It is rather likely that the innate immune response of these cells, similarly as that of other nontumor cells, controls infection by measles virus vaccine strains, which carry mutations in the V protein, a viral inhibitor of the b-interferon response (28,41). Accordingly, a low level of HSC infection with clinical measles virus isolates encoding a fully active V protein counteracting innate immunity has been previously reported (42,43).…”

Response surface methodology to determine optimal measles-specific cytokine responses in human peripheral blood mononuclear cells

“…Cells were isolated, cryopreserved and subsequently thawed for culture in RPMI-1640 culture media (Gibco, Life Technologies, Grand Island, N.Y.) containing 5% fetal calf serum (Hyclone), as we have previously described (Ryan et al, 2009; Dhiman et al, 2010). Measles virus (Edmonston vaccine strain) was grown and titered on Vero cells, as previously described (Haralambieva et al, 2010). Cytokines were quantified in cultured PBMCs (in 96-well plate format, 2 × 10 5 cells per well) following in vitro live measles virus stimulation (in triplicate) or in unstimulated cell cultures (also in triplicate), using a modified surface response methodology matrix with combinations of MOIs and incubation times as follows (Table 1): MOIs of 0.05, 0.1, 0.5, and 1 and length of cell culture of 12 h, 24h, 48h (2 days), 72h (3 days), 96h (4 days), and 120h (5 days) (Ryan et al, 2009).…”

“…Cytokines were quantified in cultured PBMCs (in 96-well plate format, 2 × 10 5 cells per well) following in vitro live measles virus stimulation (in triplicate) or in unstimulated cell cultures (also in triplicate), using a modified surface response methodology matrix with combinations of MOIs and incubation times as follows (Table 1): MOIs of 0.05, 0.1, 0.5, and 1 and length of cell culture of 12 h, 24h, 48h (2 days), 72h (3 days), 96h (4 days), and 120h (5 days) (Ryan et al, 2009). Due to limitations in the number of PBMCs, the experiment focused on a subcollection of combinations based on methodological findings from our earlier studies and the literature (Ryan et al, 2009; Dhiman et al, 2010; Haralambieva et al, 2010). Cells stimulated with 5μg/mL PHA (Sigma) were used as a positive control.…”

“…Cells stimulated with 5μg/mL PHA (Sigma) were used as a positive control. Cell-free supernatants were harvested and used for cytokine measurements by ELISA following the manufacturer’s recommendations for each kit (R&D Systems, Minneapolis, MN for IFN-λ1; Mabtech, Cincinnati, OH for IFN-α and BD Biosciences Pharmingen, San Diego, CA for the remaining cytokines) (Haralambieva et al, 2010). …”