CIITA Enhances HIV-1 Attachment to CD4+ T Cells Leading to Enhanced Infection and Cell Depletion

Porter, Kristen A.; Kelley, Lauren N.; Nekorchuk, Michael; Jones, James H.; Hahn, Amy B.; Noronha, Carlos M. C. de; Harton, Jonathan A.; Duus, Karen

doi:10.4049/jimmunol.1000830

Cited by 6 publications

(7 citation statements)

References 60 publications

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“…A). A SupT1 variant transfected with the class II transactivator (CIITA), SupT1.CIITA, has been used previously for HIV‐1 studies ; CIITA is the MHC‐II master transcriptional regulator that controls expression of MHC‐II genes, as well as processing and editing factors such as HLA‐DM and cathepsins (reviewed in ). SupT1.CIITA has been reported to express various CIITA‐controlled genes , and we observed robust expression of HLA‐DR in SupT1.CIITA cells by cell‐surface staining (Fig.…”

Section: Resultsmentioning

confidence: 99%

Naturally processed HLA‐DR3‐restricted HHV‐6B peptides are recognized broadly with polyfunctional and cytotoxic CD4 T‐cell responses

et al. 2019

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Human herpes virus 6B (HHV‐6B) is a widespread virus that infects most people early in infancy and establishes a chronic life‐long infection with periodic reactivation. CD4 T cells have been implicated in control of HHV‐6B, but antigenic targets and functional characteristics of the CD4 T‐cell response are poorly understood. We identified 25 naturally processed MHC‐II peptides, derived from six different HHV‐6B proteins, and showed that they were recognized by CD4 T‐cell responses in HLA‐matched donors. The peptides were identified by mass spectrometry after elution from HLA‐DR molecules isolated from HHV‐6B‐infected T cells. The peptides showed strong binding to matched HLA alleles and elicited recall T‐cell responses in vitro. T‐cell lines expanded in vitro were used for functional characterization of the response. Responding cells were mainly CD3+CD4+, produced IFN‐γ, TNF‐α, and low levels of IL‐2, alone or in combination, highlighting the presence of polyfunctional T cells in the overall response. Many of the responding cells mobilized CD107a, stored granzyme B, and mediated specific killing of peptide‐pulsed target cells. These results highlight a potential role for polyfunctional cytotoxic CD4 T cells in the long‐term control of HHV‐6B infection.

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Section: Resultsmentioning

confidence: 99%

Naturally processed HLA‐DR3‐restricted HHV‐6B peptides are recognized broadly with polyfunctional and cytotoxic CD4 T‐cell responses

et al. 2019

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“…The analyses and cutoff values described in this work were obtained using the system of SupT1 cells and HHV-6B strain Z29 virus. Other cell lines and virus, such as SupT1.CIITA [ 23 ], MOLT-3 and Jurkat E6 infected with HHV-6B strain Z29 and HSB-2 cells infected with HHV-6A strain GS, also showed measurable occurrence of enlarged cells, which should make them suitable for analysis by this method. However, different combinations of cell types and virus strains have specific characteristics.…”

Section: Discussionmentioning

confidence: 99%

“…Cell lines SupT1 and SupT1.CIITA [ 23 ], as well as MOLT-3 (CRL-1552™) and Jurkat E6 (TIB-152™) (both American Type Culture Collection, Manassas, VA) were infected with HHV-6B strain Z29. Cells were collected during the exponential phase of growth and incubated with the HHV-6B virus stock.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of a method to measure HHV-6B infection in vitro based on cell size

Becerra‐Artiles

Santoro

Stern

2018

Virol J

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BackgroundHuman herpesvirus 6 (HHV-6A and HHV-6B) infection of cell cultures can be measured by different methods, including immunofluorescence microscopy, flow cytometry, or quantification of virus DNA by qPCR. These methods are reliable and sensitive but require long processing times and can be costly. Another method used in the field relies on the identification of enlarged cells in the culture; this method requires little sample processing and is relatively fast. However, visual inspection of cell cultures can be subjective and it can be difficult to establish clear criteria to decide if a cell is enlarged. To overcome these issues, we explored a method to monitor HHV-6B infections based on the systematic and objective measurement of the size of cells using an imaging-based automated cell counter.ResultsThe size of cells in non-infected and HHV-6B-infected cultures was measured at different times post-infection. The relatively narrow size distribution observed for non-infected cultures contrasted with the broader distributions observed in infected cultures. The average size of cultures shifted towards higher values after infection, and the differences were significant for cultures infected with relatively high doses of virus and/or screened at longer times post-infection. Correlation analysis showed that the trend observed for average size was similar to the trend observed for two other methods to measure infection: amount of virus DNA in supernatant and the percentage of cells expressing a viral antigen. In order to determine the performance of the size-based method in differentiating non-infected and infected cells, receiver operating characteristic (ROC) curves were used to analyze the data. Analysis using size of individual cells showed a moderate performance in detecting infected cells (area under the curve (AUC) ~ 0.80-0.87), while analysis using the average size of cells showed a very good performance in detecting infected cultures (AUC ~ 0.99).ConclusionsThe size-based method proved to be useful in monitoring HHV-6B infections for cultures where a substantial fraction of cells were infected and when monitored at longer times post-infection, with the advantage of being relatively fast and easy. It is a convenient method for monitoring virus production in-vitro and bulk infection of cells.Electronic supplementary materialThe online version of this article (10.1186/s12985-017-0917-z) contains supplementary material, which is available to authorized users.

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“…CIITA-expressing HEK293T cells (6) were grown in DMEM 10% FBS, 50 M 2-mercaptoethanol, 1ϫ nonessential amino acids, 1ϫ sodium pyruvate, and fresh L-glutamine and L-histidinol.…”

Section: Methodsmentioning

confidence: 99%

Differential Transmembrane Domain GXXXG Motif Pairing Impacts Major Histocompatibility Complex (MHC) Class II Structure

Dixon

Drake

Hughes

et al. 2014

Journal of Biological Chemistry

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Background: Major histocompatibility complex class II molecules are structurally and functionally heterogeneous. Results: Combined mutagenesis and structural studies establish a role for pairing between conserved transmembrane (TM) GXXXG dimerization motifs in determining class II conformation. Conclusion: Differential pairing of highly conserved TM domain dimerization motifs contributes to class II structure and function. Significance: Global conformation contributes to the function of peptide-class II complexes.

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CIITA Enhances HIV-1 Attachment to CD4+ T Cells Leading to Enhanced Infection and Cell Depletion

Cited by 6 publications

References 60 publications

Naturally processed HLA‐DR3‐restricted HHV‐6B peptides are recognized broadly with polyfunctional and cytotoxic CD4 T‐cell responses

Naturally processed HLA‐DR3‐restricted HHV‐6B peptides are recognized broadly with polyfunctional and cytotoxic CD4 T‐cell responses

Evaluation of a method to measure HHV-6B infection in vitro based on cell size

Differential Transmembrane Domain GXXXG Motif Pairing Impacts Major Histocompatibility Complex (MHC) Class II Structure

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