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Article ; Online: Combined strategies for improving the heterologous expression of a novel xylanase from

Liu, Chun / Zhang, Yaping / Ye, Chunting / Zhao, Fengguang / Chen, Yian / Han, Shuangyan

Synthetic and systems biotechnology

2024 Volume 9, Issue 3, Page(s) 426–435

Abstract: Xylanase, an enzyme capable of hydrolyzing non-starch polysaccharides found in grain structures like wheat, has been found to improve the organizational structure of dough and thus increase its volume. In our past work, one promising xylanase FXYL ... ...

Abstract	Xylanase, an enzyme capable of hydrolyzing non-starch polysaccharides found in grain structures like wheat, has been found to improve the organizational structure of dough and thus increase its volume. In our past work, one promising xylanase FXYL derived from
Language	English
Publishing date	2024-03-26
Publishing country	China
Document type	Journal Article
ISSN	2405-805X
ISSN (online)	2405-805X
DOI	10.1016/j.synbio.2024.03.012
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: A rapid, sensitive, and reproducible in vivo PBMC humanized murine model for determining therapeutic-related cytokine release syndrome.

Ye, Chunting / Yang, Hongyuan / Cheng, Mingshan / Shultz, Leonard D / Greiner, Dale L / Brehm, Michael A / Keck, James G

FASEB journal : official publication of the Federation of American Societies for Experimental Biology

2020 Volume 34, Issue 9, Page(s) 12963–12975

Abstract: Immunotherapy is a powerful treatment strategy being applied to cancer, autoimmune diseases, allergies, and transplantation. Although therapeutic monoclonal antibodies (mAbs) have demonstrated significant clinical efficacy, there is also the potential ... ...

Abstract	Immunotherapy is a powerful treatment strategy being applied to cancer, autoimmune diseases, allergies, and transplantation. Although therapeutic monoclonal antibodies (mAbs) have demonstrated significant clinical efficacy, there is also the potential for severe adverse events, including cytokine release syndrome (CRS). CRS is characterized by the rapid production of inflammatory cytokines following delivery of therapy, with symptoms ranging from mild fever to life-threating pathology and multi-organ failure. Overall there is a paucity of models to reliably and accurately predict the induction of CRS by immune therapeutics. Here, we describe the development of a humanized mouse model based on the NOD-scid IL2rg
MeSH term(s)	Animals ; Antibodies, Monoclonal/adverse effects ; Antibodies, Monoclonal/immunology ; Cytokine Release Syndrome/chemically induced ; Cytokine Release Syndrome/immunology ; Cytokines/immunology ; Disease Models, Animal ; Female ; Leukocytes, Mononuclear/immunology ; Mice ; Mice, Inbred NOD ; Mice, SCID
Chemical Substances	Antibodies, Monoclonal ; Cytokines
Keywords	covid19
Language	English
Publishing date	2020-08-09
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	639186-2
ISSN	1530-6860 ; 0892-6638
ISSN (online)	1530-6860
ISSN	0892-6638
DOI	10.1096/fj.202001203R
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Article: Gene array analysis of PD-1H overexpressing monocytes reveals a pro-inflammatory profile

Bharaj, Preeti / Ye, Chunting / Petersen, Sean / Wang, Qianghu / Hu, Baoli / Manjunath, N / Shankar, Premlata / Yi, Guohua

Heliyon. 2018 Feb., v. 4, no. 2

2018

Abstract: We have previously reported that overexpression of Programmed Death -1 Homolog (PD-1H) in human monocytes leads to activation and spontaneous secretion of multiple pro inflammatory cytokines. Here we evaluate changes in monocytes gene expression after ... ...

Abstract	We have previously reported that overexpression of Programmed Death -1 Homolog (PD-1H) in human monocytes leads to activation and spontaneous secretion of multiple pro inflammatory cytokines. Here we evaluate changes in monocytes gene expression after enforced PD-1H expression by gene array. The results show that there are significant alterations in 51 potential candidate genes that relate to immune response, cell adhesion and metabolism. Genes corresponding to pro-inflammatory cytokines showed the highest upregulation, 7, 3.2, 3.0, 5.8, 4.4 and 3.1 fold upregulation of TNF-α, IL-1 β, IFN-α, γ, λ and IL-27 relative to vector control. The data are in agreement with cytometric bead array analysis showing induction of proinflammatory cytokines, IL-6, IL-1β and TNF-α by PD-1H. Other genes related to inflammation, include transglutaminase 2 (TG2), NF-κB (p65 and p50) and toll like receptors (TLR) 3 and 4 were upregulated 5, 4.5 and 2.5 fold, respectively. Gene set enrichment analysis (GSEA) also revealed that signaling pathways related to inflammatory response, such as NFκB, AT1R, PYK2, MAPK, RELA, TNFR1, MTOR and proteasomal degradation, were significantly upregulated in response to PD-1H overexpression. We validated the results utilizing a standard inflammatory sepsis model in humanized BLT mice, finding that PD-1H expression was highly correlated with proinflammatory cytokine production. We therefore conclude that PD-1H functions to enhance monocyte activation and the induction of a pro-inflammatory gene expression profile.
Keywords	cell adhesion ; death ; genes ; humans ; immune response ; inflammation ; interleukin-6 ; metabolism ; models ; monocytes ; protein-glutamine gamma-glutamyltransferase ; secretion ; vector control
Language	English
Dates of publication	2018-02
Publishing place	Elsevier Ltd
Document type	Article
Note	NAL-AP-2-clean
ZDB-ID	2835763-2
ISSN	2405-8440
ISSN	2405-8440
DOI	10.1016/j.heliyon.2018.e00545
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Article: A rapid, sensitive, and reproducible in vivo PBMC humanized murine model for determining therapeutic-related cytokine release syndrome

Ye, Chunting / Yang, Hongyuan / Cheng, Mingshan / Shultz, Leonard D / Greiner, Dale L / Brehm, Michael A / Keck, James G

FASEB j

Abstract	Immunotherapy is a powerful treatment strategy being applied to cancer, autoimmune diseases, allergies, and transplantation. Although therapeutic monoclonal antibodies (mAbs) have demonstrated significant clinical efficacy, there is also the potential for severe adverse events, including cytokine release syndrome (CRS). CRS is characterized by the rapid production of inflammatory cytokines following delivery of therapy, with symptoms ranging from mild fever to life-threating pathology and multi-organ failure. Overall there is a paucity of models to reliably and accurately predict the induction of CRS by immune therapeutics. Here, we describe the development of a humanized mouse model based on the NOD-scid IL2rgnull (NSG) mouse to study CRS in vivo. PBMC-engrafted NSG, NSG-MHC-DKO, and NSG-SGM3 mice were used to study cytokine release in response to treatment with mAb immunotherapies. Our data show that therapeutic-stimulated cytokine release in these PBMC-based NSG models captures the variation in cytokine release between individual donors, is drug dependent, occurs in the absence of acute xeno-GVHD, highlighting the specificity of the assay, and shows a robust response following treatment with a TGN1412 analog, a CD28 superagonist. Overall our results demonstrate that PBMC-engrafted NSG models are rapid, sensitive, and reproducible platforms to screen novel therapeutics for CRS.
Keywords	covid19
Publisher	WHO
Document type	Article
Note	WHO #Covidence: #702550
Database	COVID19

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Article: Gene array analysis of PD-1H overexpressing monocytes reveals a pro-inflammatory profile.

Bharaj, Preeti / Ye, Chunting / Petersen, Sean / Wang, Qianghu / Hu, Baoli / Manjunath, N / Shankar, Premlata / Yi, Guohua

Heliyon

2018 Volume 4, Issue 2, Page(s) e00545

Abstract	We have previously reported that overexpression of Programmed Death -1 Homolog (PD-1H) in human monocytes leads to activation and spontaneous secretion of multiple pro inflammatory cytokines. Here we evaluate changes in monocytes gene expression after enforced PD-1H expression by gene array. The results show that there are significant alterations in 51 potential candidate genes that relate to immune response, cell adhesion and metabolism. Genes corresponding to pro-inflammatory cytokines showed the highest upregulation, 7, 3.2, 3.0, 5.8, 4.4 and 3.1 fold upregulation of TNF-α, IL-1 β, IFN-α, γ, λ and IL-27 relative to vector control. The data are in agreement with cytometric bead array analysis showing induction of proinflammatory cytokines, IL-6, IL-1β and TNF-α by PD-1H. Other genes related to inflammation, include transglutaminase 2 (TG2), NF-κB (p65 and p50) and toll like receptors (TLR) 3 and 4 were upregulated 5, 4.5 and 2.5 fold, respectively. Gene set enrichment analysis (GSEA) also revealed that signaling pathways related to inflammatory response, such as NFκB, AT1R, PYK2, MAPK, RELA, TNFR1, MTOR and proteasomal degradation, were significantly upregulated in response to PD-1H overexpression. We validated the results utilizing a standard inflammatory sepsis model in humanized BLT mice, finding that PD-1H expression was highly correlated with proinflammatory cytokine production. We therefore conclude that PD-1H functions to enhance monocyte activation and the induction of a pro-inflammatory gene expression profile.
Language	English
Publishing date	2018-03-01
Publishing country	England
Document type	Journal Article
ZDB-ID	2835763-2
ISSN	2405-8440
ISSN	2405-8440
DOI	10.1016/j.heliyon.2018.e00545
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Article ; Online: Single amino acid change in gp41 region of HIV-1 alters bystander apoptosis and CD4 decline in humanized mice

Ye Chunting / Joshi Anjali / Garg Himanshu / Shankar Premlata / Manjunath N

Virology Journal, Vol 8, Iss 1, p

2011 Volume 34

Abstract: Abstract Background The mechanism by which HIV infection leads to a selective depletion of CD4 cells leading to immunodeficiency remains highly debated. Whether the loss of CD4 cells is a direct consequence of virus infection or bystander apoptosis of ... ...

Abstract	Abstract Background The mechanism by which HIV infection leads to a selective depletion of CD4 cells leading to immunodeficiency remains highly debated. Whether the loss of CD4 cells is a direct consequence of virus infection or bystander apoptosis of uninfected cells is also uncertain. Results We have addressed this issue in the humanized mouse model of HIV infection using a HIV variant with a point mutation in the gp41 region of the Env glycoprotein that alters its fusogenic activity. We demonstrate here that a single amino acid change (V38E) altering the cell-to-cell fusion activity of the Env minimizes CD4 loss in humanized mice without altering viral replication. This differential pathogenesis was associated with a lack of bystander apoptosis induction by V38E virus even in the presence of similar levels of infected cells. Interestingly, immune activation was observed with both WT and V38E infection suggesting that the two phenomena are likely not interdependent in the mouse model. Conclusions We conclude that Env fusion activity is one of the determinants of HIV pathogenesis and it may be possible to attenuate HIV by targeting gp41.
Keywords	Microbiology ; QR1-502 ; Science ; Q ; DOAJ:Microbiology ; DOAJ:Biology ; DOAJ:Biology and Life Sciences ; Medicine (General) ; R5-920 ; Medicine ; R ; DOAJ:Medicine (General) ; DOAJ:Health Sciences
Subject code	570
Language	English
Publishing date	2011-01-01T00:00:00Z
Publisher	BioMed Central
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: IL-10 exacerbates xenogeneic GVHD by inducing massive human T cell expansion.

Abraham, Sojan / Choi, Jang-gi / Ye, Chunting / Manjunath, N / Shankar, Premlata

Clinical immunology (Orlando, Fla.)

2015 Volume 156, Issue 1, Page(s) 58–64

Abstract: Although patients with GVHD have elevated serum levels of IL10, whether its role is protective or pathogenic remains unclear. Here, we used a humanized mouse model to study the role of IL-10 in GVHD. When human PBMCs were engrafted in NOD-scid IL2rγc( ... ...

Abstract	Although patients with GVHD have elevated serum levels of IL10, whether its role is protective or pathogenic remains unclear. Here, we used a humanized mouse model to study the role of IL-10 in GVHD. When human PBMCs were engrafted in NOD-scid IL2rγc(null) mice expressing human IL-10, the T cells underwent massive expansion resulting in lethality by day 21, whereas control mice survived for at least 40 days. Histopathology of the liver showed extensive mononuclear cell infiltration in IL-10 expressing but not in control mice. Corresponding to their aggressiveness, the T cells in the IL-10 group exhibited predominantly an effector memory phenotype (CD45RO(+)CD27(-)) while in control mice, the T cells were of transitional memory phenotype (CD45RO(+)CD27(+)). Further, IL-10 receptor blocking antibody was able to protect the animals from GVHD. Since our results demonstrate a direct pathogenic role for IL-10, blockade of IL-10 signaling may provide a therapeutic option for GVHD.
MeSH term(s)	Animals ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Graft vs Host Disease/physiopathology ; Heterografts ; Humans ; Interleukin-10/metabolism ; Mice ; Mice, Inbred NOD ; Models, Animal ; T-Lymphocytes/cytology
Chemical Substances	Interleukin-10 (130068-27-8)
Language	English
Publishing date	2015-01
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	1459903-x
ISSN	1521-7035 ; 1521-6616
ISSN (online)	1521-7035
ISSN	1521-6616
DOI	10.1016/j.clim.2014.11.004
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Article ; Online: Enhanced induction of HIV-specific CTL by dendritic cell-targeted delivery of SOCS-1 siRNA

Ye Chunting / Subramanya Sandesh / Kim Sang-Soo / Shankar Premalata

Retrovirology, Vol 7, Iss Suppl 1, p P

2010 Volume 12

Keywords	Medicine (General) ; R5-920 ; Medicine ; R ; DOAJ:Medicine (General) ; DOAJ:Health Sciences ; Immunologic diseases. Allergy ; RC581-607
Language	English
Publishing date	2010-05-01T00:00:00Z
Publisher	BioMed Central
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Optimizing sgRNA structure to improve CRISPR-Cas9 knockout efficiency

Dang, Ying / Jia, Gengxiang / Choi, Jennie / Ma, Hongming / Anaya, Edgar / Ye, Chunting / Shankar, Premlata / Wu, Haoquan

Genome biology. 2015 Dec., v. 16, no. 1

2015

Abstract: BACKGROUND: Single-guide RNA (sgRNA) is one of the two key components of the clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 genome-editing system. The current commonly used sgRNA structure has a shortened duplex compared with the ...

Abstract	BACKGROUND: Single-guide RNA (sgRNA) is one of the two key components of the clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 genome-editing system. The current commonly used sgRNA structure has a shortened duplex compared with the native bacterial CRISPR RNA (crRNA)–transactivating crRNA (tracrRNA) duplex and contains a continuous sequence of thymines, which is the pause signal for RNA polymerase III and thus could potentially reduce transcription efficiency. RESULTS: Here, we systematically investigate the effect of these two elements on knockout efficiency and showed that modifying the sgRNA structure by extending the duplex length and mutating the fourth thymine of the continuous sequence of thymines to cytosine or guanine significantly, and sometimes dramatically, improves knockout efficiency in cells. In addition, the optimized sgRNA structure also significantly increases the efficiency of more challenging genome-editing procedures, such as gene deletion, which is important for inducing a loss of function in non-coding genes. CONCLUSIONS: By a systematic investigation of sgRNA structure we find that extending the duplex by approximately 5 bp combined with mutating the continuous sequence of thymines at position 4 to cytosine or guanine significantly increases gene knockout efficiency in CRISPR-Cas9-based genome editing experiments.
Keywords	DNA-directed RNA polymerase ; RNA ; cytosine ; gene deletion ; gene editing ; gene targeting ; genes ; guanine ; thymine
Language	English
Dates of publication	2015-12
Size	p. 280.
Publishing place	BioMed Central
Document type	Article
ZDB-ID	2040529-7
ISSN	1474-760X ; 1465-6906
ISSN (online)	1474-760X
ISSN	1465-6906
DOI	10.1186/s13059-015-0846-3
Database	NAL-Catalogue (AGRICOLA)

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Article: Combination of IL-10 and IL-2 induces oligoclonal human CD4 T cell expansion during xenogeneic and allogeneic GVHD in humanized mice

Abraham, Sojan / Guo, Hua / Choi, Jang-gi / Ye, Chunting / Thomas, Midhun Ben / Ortega, Nora / Dwivedi, Alok / Manjunath, N / Yi, Guohua / Shankar, Premlata

Heliyon. 2017 Apr., v. 3, no. 4

2017

Abstract: IL-10 is a crucial anti-inflammatory cytokine which can also exert a seemingly divergent immunostimulatory effects under certain conditions. We found high levels of the cytokine in a xenogeneic GVHD model where NOD-scid IL2rγcnull (NSG) mice were ... ...

Abstract	IL-10 is a crucial anti-inflammatory cytokine which can also exert a seemingly divergent immunostimulatory effects under certain conditions. We found high levels of the cytokine in a xenogeneic GVHD model where NOD-scid IL2rγcnull (NSG) mice were transplanted with human PBMCs in presence of IL-2. Presence of exogenous IL-10 altered the kinetics of IL-2 induced human T cell reconstitution in vivo, showing an initial delay, followed by rapid expansion. Further, compared to IL-2 alone, treatment with IL-2 in combination with IL-10 increased survival in most animals and completely protected ∼20% of mice from GVHD. Additionally, IL-2 induced expansion of both CD4⁺ and CD8⁺ xenoreactive T cells whereas a combination of IL-2 and IL-10 resulted in selective expansion of CD4⁺ T cells only. TCR Vβ repertoire analysis of CD4⁺ T cells showed that in contrast to IL-2 alone, simultaneous presence of both cytokines drastically reduced the Vβ repertoire of the expanded CD4⁺ T cells. Highly restricted Vβ usage was also observed when the cytokine combination was tested in an allogeneic GVHD model where NOD-scid IL2rγcⁿᵘˡˡ mice expressing HLA-DR4 (NSG-DR4) were transplanted with purified CD4⁺ T cells from HLA-DR4 negative donors. Taken together, our results demonstrate that IL-10 can profoundly modulate the subset composition and repertoire of responding T cells during GVHD.
Keywords	CD4-positive T-lymphocytes ; humans ; immunostimulants ; interleukin-10 ; interleukin-2 ; models
Language	English
Dates of publication	2017-04
Publishing place	Elsevier Ltd
Document type	Article
Note	NAL-AP-2-clean
ZDB-ID	2835763-2
ISSN	2405-8440
ISSN	2405-8440
DOI	10.1016/j.heliyon.2017.e00276
Database	NAL-Catalogue (AGRICOLA)

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