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Article ; Online: Effects of blood sample storage time, temperature, anti-coagulants and blood stabiliser on lymphocyte phenotyping.

Lao, Yunyu / Quach, Alex / Perveen, Khalida / Hii, Charles / Ferrante, Antonio

2024 Volume 56, Issue 4, Page(s) 571–576

Abstract: Medical diagnostic laboratories have come under further scrutiny to ensure quality standards of their service and external quality assurance (EQA) programs involving multiple laboratories have been used to gauge this quality based on a consensus. However, ...

Abstract	Medical diagnostic laboratories have come under further scrutiny to ensure quality standards of their service and external quality assurance (EQA) programs involving multiple laboratories have been used to gauge this quality based on a consensus. However, because of the geographical distances within a country or internationally, cell surface marker expressions may change due to time delays and transport temperatures. Attention was given to this issue some decades ago and hence requires a re-evaluation in consideration of updated methods, reagents and instruments for flow cytometry and phenotyping. We have undertaken an extensive study to examine the effects of various conditions on blood storage akin to that experienced by patient samples as well as EQA programs, examining expression of lymphocyte surface markers, CD3, CD4, CD8, CD2, CD19, CD20, CD16/56 and HLA-DR. Assessment of lithium-heparin anticoagulated whole blood showed an increase in percentage of CD3
MeSH term(s)	Humans ; Temperature ; Immunophenotyping ; Flow Cytometry ; Anticoagulants/pharmacology ; Anticoagulants/therapeutic use ; Time Factors ; Lymphocytes ; Blood Preservation ; Blood Specimen Collection/methods ; Phenotype
Chemical Substances	Anticoagulants
Language	English
Publishing date	2024-02-07
Publishing country	England
Document type	Journal Article
ZDB-ID	7085-3
ISSN	1465-3931 ; 0031-3025
ISSN (online)	1465-3931
ISSN	0031-3025
DOI	10.1016/j.pathol.2023.11.011
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Article ; Online: PKCζ activation promotes maturation of cord blood T cells towards a Th1 IFN-γ propensity.

Perveen, Khalida / Quach, Alex / Stark, Michael J / Prescott, Susan / Barry, Simon C / Hii, Charles S / Ferrante, Antonio

Immunology

2023 Volume 170, Issue 3, Page(s) 359–373

Abstract: A significant number of babies present transiently with low protein kinase C zeta (PKCζ) levels in cord blood T cells (CBTC), associated with reduced ability to transition from a neonatal Th2 to a mature Th1 cytokine bias, leading to a higher risk of ... ...

Abstract	A significant number of babies present transiently with low protein kinase C zeta (PKCζ) levels in cord blood T cells (CBTC), associated with reduced ability to transition from a neonatal Th2 to a mature Th1 cytokine bias, leading to a higher risk of developing allergic sensitisation, compared to neonates whose T cells have 'normal' PKCζ levels. However, the importance of PKCζ signalling in regulating their differentiation from a Th2 to a Th1 cytokine phenotype propensity remains undefined. To define the role of PKCζ signalling in the regulation of CBTC differentiation from a Th2 to a Th1cytokine phenotype we have developed a neonatal T cell maturation model which enables the cells to develop to CD45RA
MeSH term(s)	Infant, Newborn ; Humans ; Interferon-gamma/metabolism ; Th1 Cells/metabolism ; Fetal Blood ; Cytokines/metabolism ; Cell Differentiation ; Leukocyte Common Antigens ; Th2 Cells/metabolism
Chemical Substances	Interferon-gamma (82115-62-6) ; Cytokines ; Leukocyte Common Antigens (EC 3.1.3.48)
Language	English
Publishing date	2023-06-20
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80124-0
ISSN	1365-2567 ; 0019-2805 ; 0953-4954
ISSN (online)	1365-2567
ISSN	0019-2805 ; 0953-4954
DOI	10.1111/imm.13674
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Article ; Online: Neutrophils Require Activation to Express Functional Cell-Surface Complement Receptor Immunoglobulin.

Small, Annabelle G / Perveen, Khalida / Putty, Trishni / Patel, Nikita / Quinn, Patrick / Wechalekar, Mihir D / Hii, Charles S / Quach, Alex / Ferrante, Antonio

Frontiers in immunology

2022 Volume 13, Page(s) 840510

Abstract: The phagocytosis-promoting complement receptor, Complement Receptor Immunoglobulin (CRIg), is exclusively expressed on macrophages. It has been demonstrated that expression in macrophages could be modulated by inflammatory mediators, including cytokines. ...

Abstract	The phagocytosis-promoting complement receptor, Complement Receptor Immunoglobulin (CRIg), is exclusively expressed on macrophages. It has been demonstrated that expression in macrophages could be modulated by inflammatory mediators, including cytokines. This raised the possibility that a major phagocyte, the neutrophil, may also express CRIg following activation with inflammatory mediators. Here we show that resting peripheral blood neutrophil lysates subjected to protein analysis by Western blot revealed a 35 kDa CRIg isoform, consistent with the expression of CRIg mRNA by RT-PCR. By flow cytometry, CRIg was detected intracellularly and in very minor amounts on the cell surface. Interestingly, expression on the cell surface was significantly increased to functional levels after activation with inflammatory mediators/neutrophil activators; N-Formylmethionine-leucyl-phenylalanine, tumor necrosis factor (TNF), Granulocyte-Macrophage Colony stimulating Factor (GM-CSF), bacterial lipopolysaccharide, leukotriene B4 and phorbol myristate acetate. The increase in expression required p38 MAP kinase and protein kinase C activation, as well as intracellular calcium. Neutrophils which were defective in actin microfilament reorganization due to a mutation in ARPC1B or inhibition of its upstream regulator, Rac2 lose their ability to upregulate CRIg expression. Inhibition of another small GTPase, Rab27a, with pharmacological inhibitors prevented the increase in CRIg expression, suggesting a requirement for the actin cytoskeleton and exocytosis. Engagement of CRIg on TNF-primed neutrophils with an anti-CRIg monoclonal antibody increased the release of superoxide and promoted the activation of p38 but not ERK1/ERK2 or JNK MAP kinases. The TNF-induced increase in killing of
MeSH term(s)	Cytokines/metabolism ; Granulocyte-Macrophage Colony-Stimulating Factor/metabolism ; Immunoglobulins/metabolism ; Inflammation Mediators/metabolism ; Neutrophils/metabolism ; Receptors, Complement/genetics ; Receptors, Complement/metabolism ; Tumor Necrosis Factor-alpha/metabolism
Chemical Substances	Cytokines ; Immunoglobulins ; Inflammation Mediators ; Receptors, Complement ; Tumor Necrosis Factor-alpha ; Granulocyte-Macrophage Colony-Stimulating Factor (83869-56-1)
Language	English
Publishing date	2022-03-04
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2022.840510
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Article: Predictive accuracy of anti mullerian hormone as indicator of ovarian follicle loss in cyclophosphamide treated mice.

Jamil, Zehra / Perveen, Khalida / Malik, Rabia / Avesi, Lubna

JPMA. The Journal of the Pakistan Medical Association

2017 Volume 67, Issue 10, Page(s) 1470–1475

Abstract: Objective: To evaluate the strength of anti-mullerian hormone in reflecting the stages of ovarian toxicity-induced by cyclophosphamide.: Methods: This study was conducted in December 2014 and comprised female mice that were divided into four groups: ... ...

Abstract	Objective: To evaluate the strength of anti-mullerian hormone in reflecting the stages of ovarian toxicity-induced by cyclophosphamide. Methods: This study was conducted in December 2014 and comprised female mice that were divided into four groups: group A served as control, group B received three weekly injections of cyclophosphamide, group C was co administered alpha-tocopherol along with cyclophosphamide, while group D solely received alpha-tocopherol. The ovaries were evaluated for follicular dynamics, and anti-mullerian hormone was assessed using mouse enzyme-linked immunosorbent assay kit. The data was analysed using SPSS 19. Results: There were 40 mice in the study. Histological analysis revealed severely reduced ovarian reserve in group B(p<0.01).In group C alpha-tocopherol conserved the ovarian reserve to near normal, thus follicle count was significantly higher than group B (p<0.05). However, this moderate reduction was still lower than the controls (p<0.01). Furthermore, the number of corpus lutea and atretic follicles were significantly higher in groups B and C (p<0.01). Regarding hormonal analyses in comparison to controls, anti-mullerian hormone levels were low in group B (p<0.01), while group C reported an insignificant fall in serum anti-mullerian hormone levels (p=0.101). Conclusions: There was substantial evidence that anti-mullerian hormone monitoring during chemotherapy administration may fulfil the criteria of earliest diagnostic indicator of secondary infertility.
MeSH term(s)	Animals ; Anti-Mullerian Hormone/blood ; Cyclophosphamide/adverse effects ; Female ; Mice ; Ovarian Reserve/drug effects ; Ovary/chemistry ; Ovary/drug effects ; alpha-Tocopherol/pharmacology
Chemical Substances	Anti-Mullerian Hormone (80497-65-0) ; Cyclophosphamide (8N3DW7272P) ; alpha-Tocopherol (H4N855PNZ1)
Language	English
Publishing date	2017-09-27
Publishing country	Pakistan
Document type	Journal Article
ZDB-ID	603873-6
ISSN	0030-9982
ISSN	0030-9982
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Article ; Online: Characterization of the Transient Deficiency of PKC Isozyme Levels in Immature Cord Blood T Cells and Its Connection to Anti-Allergic Cytokine Profiles of the Matured Cells.

Perveen, Khalida / Quach, Alex / Stark, Michael J / Prescott, Susan L / Barry, Simon C / Hii, Charles S / Ferrante, Antonio

International journal of molecular sciences

2021 Volume 22, Issue 23

Abstract: Cord blood T cells (CBTC) from a proportion of newborns express low/deficient levels of some protein kinase C (PKC) isozymes, with low levels of PKCζ correlating with increased risk of developing allergy and associated decrease in interferon-gamma (IFN-γ) ...

Abstract	Cord blood T cells (CBTC) from a proportion of newborns express low/deficient levels of some protein kinase C (PKC) isozymes, with low levels of PKCζ correlating with increased risk of developing allergy and associated decrease in interferon-gamma (IFN-γ) producing T cells. Interestingly, these lower levels of PKCζ were increased/normalized by supplementing women during pregnancy with n-3 polyunsaturated fatty acids. However, at present, we have little understanding of the transient nature of the deficiency in the neonate and how PKCζ relates to other PKC isozymes and whether their levels influence maturation into IFN-γ producing T cells. There is also no information on PKCζ isozyme levels in the T cell subpopulations, CD4
MeSH term(s)	CD4-Positive T-Lymphocytes/immunology ; CD8-Positive T-Lymphocytes/immunology ; Female ; Fetal Blood/cytology ; Fetal Blood/immunology ; Gestational Age ; Humans ; Infant, Newborn ; Interferon-gamma/metabolism ; Interleukin-10/metabolism ; Interleukin-2/metabolism ; Interleukin-9/metabolism ; Male ; Phytohemagglutinins/pharmacology ; Pregnancy ; Protein Kinase C/genetics ; Tumor Necrosis Factor-alpha/metabolism
Chemical Substances	IFNG protein, human ; IL10 protein, human ; IL2 protein, human ; IL9 protein, human ; Interleukin-2 ; Interleukin-9 ; Phytohemagglutinins ; TNF protein, human ; Tumor Necrosis Factor-alpha ; Interleukin-10 (130068-27-8) ; Interferon-gamma (82115-62-6) ; protein kinase C zeta (EC 2.7.11.1) ; Protein Kinase C (EC 2.7.11.13)
Language	English
Publishing date	2021-11-23
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms222312650
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Article ; Online: Cord Blood T Cells Expressing High and Low PKCζ Levels Develop into Cells with a Propensity to Display Th1 and Th9 Cytokine Profiles, Respectively.

Perveen, Khalida / Quach, Alex / McPhee, Andrew / Prescott, Susan L / Barry, Simon C / Hii, Charles S / Ferrante, Antonio

International journal of molecular sciences

2021 Volume 22, Issue 9

Abstract: Low Protein Kinase C zeta (PKCζ) levels in cord blood T cells (CBTC) have been shown to correlate with the development of allergic sensitization in childhood. However, little is known about the mechanisms responsible. We have examined the relationship ... ...

Abstract	Low Protein Kinase C zeta (PKCζ) levels in cord blood T cells (CBTC) have been shown to correlate with the development of allergic sensitization in childhood. However, little is known about the mechanisms responsible. We have examined the relationship between the expression of different levels of PKCζ in CBTC and their development into mature T cell cytokine producers that relate to allergy or anti-allergy promoting cells. Maturation of naïve CBTC was initiated with anti-CD3/-CD28 antibodies and recombinant human interleukin-2 (rhIL-2). To stimulate lymphocyte proliferation and cytokine production the cells were treated with Phytohaemagglutinin (PHA) and Phorbol myristate acetate (PMA). Irrespective of the PKCζ levels expressed, immature CBTC showed no difference in lymphocyte proliferation and the production of T helper 2 (Th2) cytokine interleukin-4 (IL-4) and Th1 cytokine, interferon-gamma (IFN-γ), and influenced neither their maturation from CD45RA
MeSH term(s)	Apoptosis ; Cell Differentiation ; Cell Proliferation ; Cell Survival ; Cytokines ; Fetal Blood/cytology ; Humans ; Protein Kinase C/metabolism ; T-Lymphocytes/enzymology ; Th1 Cells/cytology ; Th1 Cells/metabolism ; Th2 Cells/cytology ; Th2 Cells/metabolism
Chemical Substances	Cytokines ; protein kinase C zeta (EC 2.7.11.1) ; Protein Kinase C (EC 2.7.11.13)
Language	English
Publishing date	2021-05-05
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms22094907
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Article ; Online: Sticking to traditions: Anatomy education and research in Pakistan.

Shahab, Nazia / Perveen, Khalida / Hussain, Mushtaq

Anatomical sciences education

2016 Volume 9, Issue 1, Page(s) 101–102

MeSH term(s)	Anatomy/education ; Pakistan ; Research
Language	English
Publishing date	2016-01
Publishing country	United States
Document type	Letter
ZDB-ID	2483491-9
ISSN	1935-9780 ; 1935-9772
ISSN (online)	1935-9780
ISSN	1935-9772
DOI	10.1002/ase.1562
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Article: Serum anti-mullerian hormone: Correlation with the ovarian follicular dynamics in healthy mice.

Jamil, Zehra / Perveen, Khalida / Malik, Rabia / Avesi, Lubna

JPMA. The Journal of the Pakistan Medical Association

2016 Volume 66, Issue 9, Page(s) 1084–1088

Abstract: Objective: To evaluate the relationship between serum anti-Mullerian hormone and follicular dynamics in mice.: Methods: This experimental study was conducted in November, 2014 at the Dow University of Health Sciences, Karachi, and comprised ... ...

Abstract	Objective: To evaluate the relationship between serum anti-Mullerian hormone and follicular dynamics in mice. Methods: This experimental study was conducted in November, 2014 at the Dow University of Health Sciences, Karachi, and comprised laboratory-bred albino mice. They were sacrifised under anaesthesia and blood was collected via cardiac puncture to assess anti-Mullerian hormone while ovaries were collected for morphometric analyses. SPSS 19 was used for data analysis. Results: There were 20 mice with a mean weight of 25±1.89 grams, while weight of the ovaries obtained from these mice was 9.6±0.92mg. The mean serum anti-Mullerian hormone was 29.89±9.7ng/ml. On average, there were 87.8+13.54 primordial follicles, 51.85±8.36 primary, 20.35±5.57 secondary, 11.30±3.38 early antral and 3.05 ± 1.27 late antral follciles (p<0.001; p=0.06).. Conclusions: Association of anti-Mullerian hormone with follicle dynamics reflected its role as a true ovarian reserve marker. Its assessment was of great significance in infertile women as well as young patients receiving chemotherapy.
MeSH term(s)	Animals ; Anti-Mullerian Hormone/blood ; Anti-Mullerian Hormone/metabolism ; Biomarkers ; Female ; Humans ; Infertility, Female/blood ; Mice ; Ovarian Follicle/metabolism ; Ovary
Chemical Substances	Biomarkers ; Anti-Mullerian Hormone (80497-65-0)
Language	English
Publishing date	2016-09-21
Publishing country	Pakistan
Document type	Journal Article
ZDB-ID	603873-6
ISSN	0030-9982
ISSN	0030-9982
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Article ; Online: Validation of monoclonal anti-PKC isozyme antibodies for flow cytometry analyses in human T cell subsets and expression in cord blood T cells.

Perveen, Khalida / Quach, Alex / McPhee, Andrew / Prescott, Susan L / Barry, Simon C / Hii, Charles S / Ferrante, Antonio

Scientific reports

2019 Volume 9, Issue 1, Page(s) 9263

Abstract: T cells from neonates (cord blood) with a tendency to develop allergic diseases express low PKCζ levels. More extensive investigations into PKC isozyme levels in T cell subsets and changes during neonatal T cell maturation are hampered by limitations of ... ...

Abstract	T cells from neonates (cord blood) with a tendency to develop allergic diseases express low PKCζ levels. More extensive investigations into PKC isozyme levels in T cell subsets and changes during neonatal T cell maturation are hampered by limitations of Western blot analyses. We have undertaken to validating the specificity of commercially available antibodies marketed for flow cytometry to measure PKCα, βI, βII, δ, ε, η, θ, ζ, ι/λ and μ. Western blot analyses of human peripheral blood mononuclear cell (PBMC) lysates demonstrated that some antibodies were unsuitable for flow cytometry assays. A panel of antibodies with the desirable specificity and reliability in the flow cytometry assay were identified using both PBMC and whole blood assays. The results showed that all PKC isozymes were expressed in CD4
MeSH term(s)	Animals ; Antibodies, Monoclonal/blood ; Antibodies, Monoclonal/immunology ; Cell Differentiation ; Cells, Cultured ; Fetal Blood/immunology ; Fetal Blood/metabolism ; Flow Cytometry/methods ; Humans ; Isoenzymes ; Leukocytes, Mononuclear/immunology ; Leukocytes, Mononuclear/metabolism ; Mice ; Protein Kinase C/antagonists & inhibitors ; Protein Kinase C/immunology ; T-Lymphocyte Subsets/immunology
Chemical Substances	Antibodies, Monoclonal ; Isoenzymes ; Protein Kinase C (EC 2.7.11.13)
Language	English
Publishing date	2019-06-25
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Validation Study
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-019-45507-2
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Article ; Online: Human Dendritic Cells Express the Complement Receptor Immunoglobulin Which Regulates T Cell Responses.

Munawara, Usma / Perveen, Khalida / Small, Annabelle G / Putty, Trishni / Quach, Alex / Gorgani, Nick N / Hii, Charles S / Abbott, Catherine A / Ferrante, Antonio

Frontiers in immunology

2019 Volume 10, Page(s) 2892

Abstract: The B7 family-related protein V-set and Ig containing 4 (VSIG4), also known as Z39Ig and Complement Immunoglobulin Receptor (CRIg), is the most recent of the complement receptors to be identified, with substantially distinct properties from the classical ...

Abstract	The B7 family-related protein V-set and Ig containing 4 (VSIG4), also known as Z39Ig and Complement Immunoglobulin Receptor (CRIg), is the most recent of the complement receptors to be identified, with substantially distinct properties from the classical complement receptors. The receptor displays both phagocytosis-promoting and anti-inflammatory properties. The receptor has been reported to be exclusively expressed in macrophages. We now present evidence, that CRIg is also expressed in human monocyte-derived dendritic cells (MDDC), including on the cell surface, implicating its role in adaptive immunity. Three CRIg transcripts were detected and by Western blotting analysis both the known Long (L) and Short (S) forms were prominent but we also identified another form running between these two. Cytokines regulated the expression of CRIg on dendritic cells, leading to its up- or down regulation. Furthermore, the steroid dexamethasone markedly upregulated CRIg expression, and in co-culture experiments, the dexamethasone conditioned dendritic cells caused significant inhibition of the phytohemagglutinin-induced and alloantigen-induced T cell proliferation responses. In the alloantigen-induced response the production of IFNγ, TNF-α, IL-13, IL-4, and TGF-β1, were also significantly reduced in cultures with dexamethasone-treated DCs. Under these conditions dexamethasone conditioned DCs did not increase the percentage of regulatory T cells (Treg). Interestingly, this suppression could be overcome by the addition of an anti-CRIg monoclonal antibody to the cultures. Thus, CRIg expression may be a control point in dendritic cell function through which drugs and inflammatory mediators may exert their tolerogenic- or immunogenic-promoting effects on dendritic cells.
MeSH term(s)	Biomarkers ; Coculture Techniques ; Cytokines/metabolism ; Dendritic Cells/immunology ; Dendritic Cells/metabolism ; Gene Expression Regulation ; Humans ; Immunity, Cellular/genetics ; Immunomodulation ; Immunophenotyping ; Receptors, Complement/genetics ; Receptors, Complement/metabolism ; T-Lymphocyte Subsets/immunology ; T-Lymphocyte Subsets/metabolism ; T-Lymphocytes/immunology ; T-Lymphocytes/metabolism
Chemical Substances	Biomarkers ; Cytokines ; Receptors, Complement
Language	English
Publishing date	2019-12-10
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2019.02892
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