Cytokine Release in HR-HPV(+) Women without and with Cervical Dysplasia (CIN II and III) or Carcinoma, Compared with HR-HPV(−) Controls

Aims. We investigated the effect of HR-HPV infection on the capacity of the cytokine network in whole blood cultures during carcinogenesis of cervical carcinoma.Methods. Thirty-nine women with moderate dysplasia, severe dysplasia, cervical carcinoma, or without dysplasia formed the study group. The control group consisted of 10 HR-HPV-negative women without CIN. Whole blood cultures were stimulated with phytohemagglutinin (PHA) and concentrations of tumour necrosis factorα(TNFα), interferonγ(IFNγ), interleukin 2 (IL-2), interleukin 12 (IL-12), interleukin 4 (IL-4), and interleukin 10 (IL-10) were determined by ELISAs.Results. A significant increase in cytokine release was detected in HR-HPV-positive women without dysplasia. In women with cervical cancer, release of IFNγand IL-12 was of the same magnitude as in HR-HPV-positive women without clinical manifestations. Most Th1-type/Th2-type ratios decreased form CIN II to CIN III, and increased from CIN III to invasive carcinoma.Conclusions. (1) Infection with HR-HPV without expression of cervical dysplasia induces activation of the cytokine network. (2) Increases in ratios of Th1-type to Th2-type cytokines at the stage of cervical carcinoma were found by comparison with stage CIN III. (3) Significant changes in the kinetics of cytokine release to a Th2-type immune response in blood of women with cervical dysplasia occurred progressively from CIN II to CIN III.

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Interleukin-12 (IL-12) Production in Whole Blood Cultures From Human Immunodeficiency Virus-Infected Individuals Studied in Relation to IL-10 and Prostaglandin E2 Production

Blood ◽

10.1182/blood.v89.2.570 ◽

1997 ◽

Vol 89 (2) ◽

pp. 570-576 ◽

Cited By ~ 29

Author(s):

Linde Meyaard ◽

Egbert Hovenkamp ◽

Nadine Pakker ◽

Tineke C.T.M. van der Pouw Kraan ◽

Frank Miedema

Keyword(s):

Human Immunodeficiency Virus ◽

T Cell ◽

Prostaglandin E2 ◽

Whole Blood ◽

Peripheral Blood ◽

Blood Cultures ◽

Pge2 Production ◽

Interleukin 12 ◽

Immunodeficiency Virus

Abstract The role of interleukin-12 (IL-12) in Th1 cell differentiation is well established. The heterodimer p70, composed of a p40 and a p35 chain, is the biologically active form. IL-12 production by human monocytes is enhanced by interferon-γ (IFN-γ) and inhibited by IL-10 and prostaglandin E2 (PGE2 ). Peripheral blood mononuclear cells from human immunodeficiency virus (HIV)-infected individuals reportedly have impaired IL-12 p40 and p70 production on stimulation with Staphylococcus aureus Cowan I (SAC) in vitro. Both PGE2 and IL-10 previously were proposed to be instrumental in this defect in IL-12 production. Here, we studied IL-12 p40 and p70 production in relation to IL-10 and PGE2 production in whole blood cultures from HIV-infected individuals. On stimulation with lipopolysaccharide, IL-12 production was normal. However, on stimulation with SAC, IL-12 p40 and p70 production was decreased in HIV-infected individuals and correlated significantly with decreased peripheral blood CD4+ T-cell number and T-cell reactivity to CD3 monoclonal antibody in vitro. However, IL-10 and PGE2 production in cultures from HIV-infected individuals was normal and did not relate to IL-12 production. In conclusion, IL-12 production by cells from HIV-infected individuals is impaired under certain conditions in vitro and this decrease is independent of IL-10 or PGE2 production.

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Downregulation of proinflammatory cytokine release in whole blood from septic patients

Blood ◽

10.1182/blood.v85.5.1341.bloodjournal8551341 ◽

1995 ◽

Vol 85 (5) ◽

pp. 1341-1347 ◽

Cited By ~ 326

Author(s):

W Ertel ◽

JP Kremer ◽

J Kenney ◽

U Steckholzer ◽

D Jarrar ◽

...

Keyword(s):

Severe Sepsis ◽

Mrna Expression ◽

Proinflammatory Cytokines ◽

Whole Blood ◽

Mononuclear Cells ◽

Natural Infection ◽

Tnf Alpha ◽

Control Group ◽

Cytokine Release ◽

Septic Group

Using animal models or healthy volunteers, injection of lipopolysaccharide (LPS) or bacteria causes activation of macrophages with excessive synthesis and secretion of proinflammatory cytokines. Although these models mimic the effects of LPS in the host, they may represent more of an experimental expression of endotoxemia than natural infection itself. Therefore, as an ex vivo model of sepsis, whole blood from 15 patients with severe sepsis and 20 control patients without infection was stimulated with LPS to study the kinetics of mRNA expression and release of proinflammatory cytokines, tumor necrosis factor (TNF)-alpha, interleukin (IL)-1 beta, and IL-6. Stimulation of whole blood with 1 microgram/mL LPS resulted in a maximum increase of cytokine secretion in the control group, while a marked (P < .01) depression of TNF-alpha, IL-1 beta, and IL-6 release was observed in the septic group, which persisted up to 10 days after study enrollment. While IL-1 beta mRNA expression was similar in peripheral blood mononuclear cells (PBMCs) harvested from LPS-stimulated whole blood in septic and control patients, the half-life and consequently the expression of TNF-alpha and IL-6 mRNA were strongly reduced in the septic group. These data indicate a downregulatory mechanism of cytokine release in whole blood from patients with severe sepsis that occurs on different levels. Although excessive secretion of proinflammatory cytokines has been considered deleterious for the host, the reduced capacity of PBMCs in whole blood from septic patients to synthesize and secrete proinflammatory cytokines to an inflammatory stimulus may result in immunodeficiency, because these cytokines in low concentrations are involved in the upregulation of essential cellular and humoral immune functions.

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Altered Salivary Protein Profiles among Individuals Diagnosed with Cervical Dysplasia and Cervical Carcinoma in Situ

Cancer and Clinical Oncology ◽

10.5539/cco.v5n1p63 ◽

2016 ◽

Vol 5 (1) ◽

pp. 63 ◽

Cited By ~ 1

Author(s):

Charles Streckfus

Keyword(s):

Cervical Cancer ◽

Cancer Progression ◽

Cervical Carcinoma ◽

Carcinoma In Situ ◽

Cervical Dysplasia ◽

Control Group ◽

Whole Saliva ◽

Healthy Control ◽

By Products

Objective: The objective of this study was to determine if protein-by-products secondary to cervical cancer oncogenes appear in the saliva.Methods: Four pooled (n=10 subjects/pool) stimulated whole saliva specimens from women were analyzed. One pooled specimen was from healthy women, while other pooled specimens were from women diagnosed with CIN 2 moderate cervical dysplasia (n=10), CIN 3 severe cervical dysplasia (n=10) and the other pooled group from women diagnosed with cervical carcinoma in situ (n=10). Differential expression of proteins was measured by isotopically tagging proteins in each groups and comparing them to the healthy control group. Saliva from each of the pooled samples was trypsinized and the peptide digests labeled with the appropriate iTRAQ reagent. Labeled peptides from each of the digests were combined and analyzed by reverse phase (C18) capillary chromatography on an Applied Biosystems QStar LC-MS/MS mass spectrometer equipped with an LC-Packings HPLC.Results: The results of the salivary analyses yielded approximately 133 proteins in the saliva specimens. Forty eight proteins were differentially expressed between the healthy control pool, precancerous and cancerous conditions.Conclusions: The study suggests that saliva is a fluid suffused with solubilized by-products of oncogenic expression and that these proteins may be modulated secondary to precancerous and cancerous conditions and that these proteins may be useful in the study of cervical cancer progression, treatment efficacy and the tailoring of individualized patient care.

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Sample Aging Profoundly Reduces Monocyte Responses in Human Whole Blood Cultures

Journal of Immunology Research ◽

10.1155/2018/8901485 ◽

2018 ◽

Vol 2018 ◽

pp. 1-9

Author(s):

H. W. Grievink ◽

M. Moerland

Keyword(s):

Whole Blood ◽

Intracellular Cytokine Staining ◽

Drug Effects ◽

Blood Cultures ◽

Blood Collection ◽

Innate Immune Responses ◽

Cytokine Release ◽

Human Whole Blood ◽

The Public

Human whole blood cultures are widely used for the investigation of physiological pathways and drug effectsin vitro. Detailed information on the effect of “sample aging” (the time span between blood collection and experimental start) on the experimental outcome is not readily available in the public domain. We studied the effect of sample aging on the ability of immune cells to respond to cell-specific immune triggers (LPS, PMA/ionomycin, and SEB). Sample aging at room temperature profoundly inhibited the LPS-induced monocytic cytokine release in minimally diluted whole blood cultures. The reduction ranged from 20–50% after 30 minutes to 80–100% after 10 hours and differed between cytokines (IL-1β, IL-2, IL-6, IFNγ, and TNFα). Sample storage at 4°C or 37°C even worsened this. PMA/ionomycin- and SEB-induced cytokine release, both mainly T-cell-driven, were also reduced by sample aging but to a lesser extent (20–50% after 24 hours). Intracellular cytokine staining revealed that the number of LPS-responding cells was not impacted by sample aging and reduced LPS responsivity could also not be explained by apoptosis or downregulated TLR4 expression. Thus, we speculate that sample aging induces an inhibitory pathway downstream from TLR4 in monocytes. These results underline the importance of quick sample handling when investigating innate immune responses in whole blood, especially for monocyte responses.

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Viscosity and Red Cell Deformity in Arterial Disease

10.1055/s-0038-1665789 ◽

1979 ◽

Author(s):

G Cella ◽

H de Haas ◽

M Rampling ◽

V Kakkar

Keyword(s):

Vascular Disease ◽

Blood Viscosity ◽

Whole Blood ◽

Arterial Disease ◽

Plasma Viscosity ◽

Control Group ◽

Fibrinogen Concentration ◽

Red Cell ◽

Whole Blood Viscosity ◽

Significant Difference

Haemorrheological factors have been shown to be affected in many kings of vascular disease. The present study was undertaken to correlate these factors in normal subjects and patients suffering from peripheral arterial disease. Twenty-two patients were investigated; they had moderate or severe intermittent claudication, extent of disease being confirmed by aorto-arteriography and ankle-systolic pressure studies. Twenty-five controls with no symptoms or signs of arterial disease were selected with comparable age and sex distribution. Whole blood viscosity was measured at shear rates of 230 secs-1 and 23 secs-lat 37°c using a Wells Brookfield cone plate microvisco meter. Plasma viscosity was also measured in an identical manner. Erythrocyte flexibility was measured by centrifuge technique and fibrinogen concentration as well as haematocrit by standard techniques. The fibrinogen concentration appeared to be the only significant parameter; the mean concentration in patients with peripheral vascular disease of 463 ± 73mg/l00ml in the control group ( < 0.05). Although whole blood viscosity was high in patients, when corrected to a common haematocrit, there was no significant difference between patients and controls. The same megative correlation was found for plasma viscosity. The red cell flexibility was found to be increased in patients as compared to the control group, but this effect appeared to be simply proportional to the fibrinogen concentration.

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Cytokine profiles and Lactobacillus species presence in pre-menopausal subjects with genital Mycoplasma genitalium or Ureaplasma urealyticum colonization

Women s Health ◽

10.1177/17455065211009181 ◽

2021 ◽

Vol 17 ◽

pp. 174550652110091

Author(s):

John Garza ◽

Kushal Gandhi ◽

Sarah Choi ◽

Asley Sanchez ◽

Gary Ventolini

Keyword(s):

Ureaplasma Urealyticum ◽

Mycoplasma Genitalium ◽

Interleukin 12 ◽

Lactobacillus Species ◽

Control Group ◽

Study Group ◽

Species Dominance ◽

Cytokine Profiles ◽

Menopausal Women ◽

Significant Difference

Background and Purpose: Lactobacilli play a vital role in protecting the vagina against pathogens. Cytokines are vital components of defense against infections in women. The genital mycoplasmas, Mycoplasma genitalium and Ureaplasma urealyticum, are associated with various infectious diseases in adults and infants. The objective of our study is to identify differences in cytokine profile and Lactobacillus species dominance between a study group of non-pregnant pre-menopausal women with genital M. genitalium or U. urealyticum colonization and a control group of non-pregnant pre-menopausal women without genital M. genitalium or U. urealyticum colonization. Methods: A real-time polymerase chain reaction was performed to measure Lactobacillus species in vaginal swab samples. Cytokine analysis was performed using multiplex immunoassay techniques. Analysis of variance confirmed a significant difference in cytokine profiles between patient groups, with t-tests identifying the most significantly different cytokines. Categorical data analysis identified significant patterns of relative Lactobacillus species dominance in the study group. Results: Lactobacillus iners was the predominant Lactobacillus species in the control group ( p = 0.005). There were no dominant Lactobacillus species observed in the study group. Vascular endothelial growth factor A ( p = 0.002), interleukin-8 ( p = 0.001), and interleukin-1β ( p = 0.049) were expressed significantly higher in the study group, whereas interleukin-1 receptor antagonist ( p < 0.001), interleukin-10 ( p = 0.001), interleukin-12 ( p = 0.002), and interferon-γ ( p = 0.022) were expressed higher in the control group. Association matrices for cytokines were significantly different between two groups ( p < 0.001), with mostly negative associations in the control group and mostly positive associations in the study group. Conclusion: Cytokine levels, their associations, and the patterns of Lactobacillus species dominance are observed to significantly diverge on the basis of M. genitalium and U. urealyticum colonization among non-pregnant pre-menopausal women.

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Characterization of Feline Whole-blood Cultures and Determination of the Frequency of Radiation-induced Dicentrics in Human and Feline Lymphocytes

International Journal of Radiation Biology and Related Studies in Physics Chemistry and Medicine ◽

10.1080/09553007914550421 ◽

1979 ◽

Vol 35 (4) ◽

pp. 351-359 ◽

Cited By ~ 14

Author(s):

G. Stephan ◽

I.-D. Adler ◽

D. Schwartz-Porsche ◽

K.-U. Hollihn ◽

G. Obe

Keyword(s):

Whole Blood ◽

Blood Cultures ◽

Radiation Induced

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Identification of Cytokines in Whole Blood for Differential Diagnosis of Tuberculosis versus Pneumonia

Clinical and Vaccine Immunology ◽

10.1128/cvi.00526-09 ◽

2010 ◽

Vol 17 (5) ◽

pp. 771-777 ◽

Cited By ~ 5

Author(s):

Wen-Lin Su ◽

Wann-Cherng Perng ◽

Ching-Hui Huang ◽

Cheng-Yu Yang ◽

Chin-Pyng Wu ◽

...

Keyword(s):

Whole Blood ◽

Blood Cells ◽

Cutoff Point ◽

Interleukin 12 ◽

Operating Characteristics ◽

Percent Change ◽

Ifn Γ ◽

Sensitivity Specificity ◽

Whole Blood Cells ◽

Stimulation Of

ABSTRACT Differentiating tuberculosis (TB) from pneumonia remains a challenge. We evaluated the cytokine profiles of whole blood cells from patients with TB (n = 38) or pneumonia (n = 30) and from healthy individuals (n = 30) before and after stimulating cells with ESAT-6 or lipopolysaccharide (LPS). When the percent change in the levels of gamma interferon (IFN-γ) after stimulation with ESAT-6 was used in receiver operating characteristics (ROC) analysis (a graphic method to determine the diagnostic accuracy of a test) to identify a patient with TB, the area under the curve (AUC) was 90.4%, and a cutoff point of a 3.59% change produced a corresponding sensitivity, specificity, and accuracy of over 80%. When the change in IFN-γ after stimulation of blood cells with LPS was used to identify a patient with pneumonia, the AUC reached 89.1%, and a cutoff point of 3.59% produced a sensitivity, specificity, and accuracy of approximately 80% each. When the change in interleukin-12 (IL-12) after stimulation of blood cells with LPS was selected to define a patient with pneumonia, the AUC was 85.2%, and a cutoff point of 2.08% gave a sensitivity, specificity, and accuracy of 80.0%, 78.9%, and 79.4%, respectively. We conclude that the percent change in IFN-γ after stimulation of whole blood cells with ESAT-6 may differentiate patients with TB from patients with pneumonia. The percent change in IFN-γ and IL-12 after LPS stimulation of whole blood cells could differentiate patients with pneumonia from patients with TB.

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The 795CT polymorphism in osteopontin gene is not associated with multiple sclerosis in a Spanish population

Multiple Sclerosis Journal ◽

10.1177/1352458506070944 ◽

2007 ◽

Vol 13 (2) ◽

pp. 250-252 ◽

Cited By ~ 6

Author(s):

A Mas ◽

A Martínez ◽

V De Las Heras ◽

M Bartolomé ◽

Eg De La Concha ◽

...

Keyword(s):

Multiple Sclerosis ◽

Low Frequency ◽

Interleukin 10 ◽

Interleukin 12 ◽

Italian Population ◽

Cytokine Network ◽

Activated T Cells ◽

Negative Finding ◽

The Central Nervous System ◽

Ms Pathogenesis

Multiple sclerosis (MS) is an inflammatory disease affecting the central nervous system. The dysregulation of the cytokine network is an important component of its pathogenesis. One of the cytokines produced by activated T-cells is osteopontin (OPN). OPN enhances the production of the pro-inflammatory cytokines, interleukin-12 and interferon-gamma, while reducing interleukin-10 levels. Therefore, OPN is considered a pro-inflammatory cytokine, and could play a key role in MS pathogenesis. The OPN gene contains several common polymorphisms, distributed in two main haplotypes, which may modulate its production or activity. A total of 326 MS patients and 484 healthy controls were typed for 795CT OPN polymorphism. In order to perform a familial study, 51 progenitor pairs were also included. No difference was found in the case-control or family study. This negative finding is inconsistent with a previous haplotype study in an Italian population, where the haplotype associated carried the low-frequency allele in position 795. In a Japanese population, a similar study yielded no association with this polymorphism. In conclusion, our data suggest that the 795 polymorphism does not play an etiological role per se and the haplotype structure may differ from one population to another. Multiple Sclerosis 2007; 13: 250–252. http://msj.sagepub.com

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