Intracellular Cytokine Profile of Cord and Adult Blood Lymphocytes

Blood ◽  
1998 ◽  
Vol 92 (1) ◽  
pp. 11-18 ◽  
Author(s):  
I.M.H. Chalmers ◽  
G. Janossy ◽  
M. Contreras ◽  
C. Navarrete
Keyword(s):  
Flow Cytometry ◽  
Cord Blood ◽  
Mononuclear Cells ◽  
Interleukin 2 ◽  
Peripheral Blood Lymphocytes ◽  
Cytokine Profile ◽  
Color Flow ◽  
Blood Lymphocytes ◽  
Tnf Α ◽  
Ifn Γ

Umbilical cord blood (CB) transplantation is thought to be associated with a reduced risk of severe graft-versus-host-disease (GVHD) compared with bone marrow transplantation (BMT). The cytokine cascade is known to be important in the pathogenesis of GVHD; however, previous studies investigating the cytokine secretion pattern of CB cells have been contradictory because of variations in experimental techniques. In this study, the cytokine profile of cord and adult blood lymphocytes and lymphocyte subsets has been assessed at the single-cell level by flow cytometry, using CD4/CD8 and CD45RA/CD45RO markers. Cord and adult blood mononuclear cells were stimulated with phorbol 12-myristate 13-acetate (PMA) and ionomycin in the presence of monensin. After 4 to 24 hours of incubation, interleukin-2 (IL-2), IL-4, interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α) production was measured by three-color flow cytometry. The results show that cord blood lymphocytes (CBL) produce less IL-2, IL-4, IFN-γ, and TNF-α than adult peripheral blood lymphocytes (ABL). Further subset analysis showed that in CBL the majority of cytokine producing cells were CD4+CD45RA+, whereas in ABL the cytokine-producing cells were both CD4+CD45RO+ and CD8+CD45RO+. These results suggest that the reduced incidence of GVHD in CB transplantation may partly due to the altered cytokine profile seen in CBL.

Expression and coreceptor activity of STRL33/Bonzo on primary peripheral blood lymphocytes

Blood ◽  
2000 ◽  
Vol 96 (1) ◽  
pp. 41-49 ◽  
Author(s):  
Matthew Sharron ◽  
Stefan Pöhlmann ◽  
Ken Price ◽  
Elias Lolis ◽  
Monica Tsang ◽  
...  
Keyword(s):  
Rhesus Macaque ◽  
Expression Pattern ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Interleukin 2 ◽  
Peripheral Blood Lymphocytes ◽  
Blood Lymphocytes ◽  
Immunodeficiency Virus ◽  
Hiv 1

Abstract CCR5 and CXCR4 are the major coreceptors that mediate human immunodeficiency virus 1 (HIV-1) infection, while most simian immunodeficiency virus (SIV) isolates use CCR5. A number of alternative coreceptors can also mediate infection of some virus strains in vitro, although little is known about their in vivo relevance. Therefore, we characterized the expression pattern and coreceptor activity of one of these alternative coreceptors, STRL33/Bonzo, using a newly developed monoclonal antibody. In addition to being highly expressed (approximately 1000-7000 STRL33 ABS [antibody binding sites]) on specific subsets of natural killer cells (CD3−/CD16−/low/CD56+ and CD3−/CD16low/CD56−) and CD19+ B lymphocytes (approximately 300-5000 STRL33 ABS), STRL33 was expressed at levels sufficient to support virus infection on freshly isolated, truly naive CD4+/CD45RA+/CD62L+cells (6000-11 000 ABS). STRL33 expression on peripheral blood mononuclear cells (PBMCs) was increased by mitogenic stimulation (OKT3/IL-2 [interleukin-2] had a greater effect than phytohemaglutinin (PHA)/IL-2), but it was dramatically decreased upon Ficoll purification. Infection of CCR5− human peripheral blood lymphocytes (PBLs) showed that 2 different SIV envelope (Env) proteins mediated entry into STRL33+cells. More importantly, the preferential infection of STRL33+ cells in CCR5− PBLs by an R5/X4/STRL33 HIV-1 maternal isolate in the presence of a potent CXCR4 antagonist (AMD3100) suggests that STRL33 can be used as a coreceptor by HIV-1 on primary cells. Rhesus macaque (rh) STRL33 was used less efficiently than human STRL33 by the majority of SIV Env proteins tested despite similar levels of expression, thereby making it less likely that STRL33 is a relevant coreceptor in the rhesus macaque system. In summary, the expression pattern and coreceptor activity of STRL33 suggest its involvement in trafficking of tumor-infiltrating lymphocytes and indicate that STRL33 may be a relevant coreceptor in vivo.

Expression and coreceptor activity of STRL33/Bonzo on primary peripheral blood lymphocytes

Blood ◽  
2000 ◽  
Vol 96 (1) ◽  
pp. 41-49 ◽  
Author(s):  
Matthew Sharron ◽  
Stefan Pöhlmann ◽  
Ken Price ◽  
Elias Lolis ◽  
Monica Tsang ◽  
...  
Keyword(s):  
Rhesus Macaque ◽  
Expression Pattern ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Interleukin 2 ◽  
Peripheral Blood Lymphocytes ◽  
Blood Lymphocytes ◽  
Immunodeficiency Virus ◽  
Hiv 1

CCR5 and CXCR4 are the major coreceptors that mediate human immunodeficiency virus 1 (HIV-1) infection, while most simian immunodeficiency virus (SIV) isolates use CCR5. A number of alternative coreceptors can also mediate infection of some virus strains in vitro, although little is known about their in vivo relevance. Therefore, we characterized the expression pattern and coreceptor activity of one of these alternative coreceptors, STRL33/Bonzo, using a newly developed monoclonal antibody. In addition to being highly expressed (approximately 1000-7000 STRL33 ABS [antibody binding sites]) on specific subsets of natural killer cells (CD3−/CD16−/low/CD56+ and CD3−/CD16low/CD56−) and CD19+ B lymphocytes (approximately 300-5000 STRL33 ABS), STRL33 was expressed at levels sufficient to support virus infection on freshly isolated, truly naive CD4+/CD45RA+/CD62L+cells (6000-11 000 ABS). STRL33 expression on peripheral blood mononuclear cells (PBMCs) was increased by mitogenic stimulation (OKT3/IL-2 [interleukin-2] had a greater effect than phytohemaglutinin (PHA)/IL-2), but it was dramatically decreased upon Ficoll purification. Infection of CCR5− human peripheral blood lymphocytes (PBLs) showed that 2 different SIV envelope (Env) proteins mediated entry into STRL33+cells. More importantly, the preferential infection of STRL33+ cells in CCR5− PBLs by an R5/X4/STRL33 HIV-1 maternal isolate in the presence of a potent CXCR4 antagonist (AMD3100) suggests that STRL33 can be used as a coreceptor by HIV-1 on primary cells. Rhesus macaque (rh) STRL33 was used less efficiently than human STRL33 by the majority of SIV Env proteins tested despite similar levels of expression, thereby making it less likely that STRL33 is a relevant coreceptor in the rhesus macaque system. In summary, the expression pattern and coreceptor activity of STRL33 suggest its involvement in trafficking of tumor-infiltrating lymphocytes and indicate that STRL33 may be a relevant coreceptor in vivo.

scholarly journals Dimethyl Sulfoxide (DMSO) Decreases Cell Proliferation and TNF-α, IFN-γ, and IL-2 Cytokines Production in Cultures of Peripheral Blood Lymphocytes

Molecules ◽  
2017 ◽  
Vol 22 (11) ◽  
pp. 1789 ◽  
Author(s):  
Lucas de Abreu Costa ◽  
Marcelo Henrique Fernandes Ottoni ◽  
Michaelle dos Santos ◽  
Agnes Meireles ◽  
Valéria Gomes de Almeida ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Dimethyl Sulfoxide ◽  
Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Blood Lymphocytes ◽  
Tnf Α ◽  
Ifn Γ

scholarly journals Cytokine Profiles for Peripheral Blood Lymphocytes from Patients with Active Pulmonary Tuberculosis and Healthy Household Contacts in Response to the 30-Kilodalton Antigen ofMycobacterium tuberculosis

1998 ◽  
Vol 66 (1) ◽  
pp. 176-180 ◽  
Author(s):  
Martha Torres ◽  
Teresa Herrera ◽  
Hector Villareal ◽  
Elizabeth A. Rich ◽  
Eduardo Sada
Keyword(s):  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Interleukin 2 ◽  
Peripheral Blood Lymphocytes ◽  
Peripheral Blood Mononuclear ◽  
Active Pulmonary Tuberculosis ◽  
Household Contacts ◽  
Reverse Transcription Pcr ◽  
Ifn Γ ◽  
Cellular Immune

ABSTRACT Patients with active tuberculosis (TB) have a stronger humoral but a poorer cellular immune response to the secreted 30-kDa antigen (Ag) of Mycobacterium tuberculosis than do healthy household contacts (HHC), who presumably are more protected against disease. The basis for this observation was studied by examining the Th1 (interleukin 2 [IL-2] and gamma interferon [IFN-γ])- and Th2 (IL-10 and IL-4)-type cytokines produced in response to the 30-kDa Ag by peripheral blood mononuclear cells (PBMC) from patients with active pulmonary TB (n = 7) and from HHC who were tuberculin (purified protein derivative) skin test positive (n = 12). Thirty-kilodalton-Ag-stimulated PBMC from TB patients produced significantly lower levels of IFN-γ (none detectable) than did those from HHC (212 ± 73 pg/ml, mean ± standard error) (P < 0.001). Likewise, 30-kDa-Ag-stimulated PBMC from TB patients failed to express IFN-γ mRNA by reverse transcription-PCR, whereas cells from HHC expressed the IFN-γ gene. In contrast, 30-kDa-Ag-stimulated PBMC from TB patients produced significantly higher levels of IL-10 (403 ± 80 pg/ml) than did those from HHC (187 ± 66 pg/ml) (P < 0.013), although cells from both groups expressed the IL-10 gene. IL-2 and IL-4 were not consistently produced, and their genes were not expressed by 30-kDa-Ag-stimulated cells from either TB patients or HHC. After treatment with antituberculous drugs, lymphocytes from four of the seven TB patients proliferated and three of them expressed IFN-γ mRNA in response to the 30-kDa Ag and produced decreased levels of IL-10.

scholarly journals Expression of the interleukin-2 receptor gamma chain on cord blood mononuclear cells

Blood ◽  
1996 ◽  
Vol 87 (8) ◽  
pp. 3344-3350 ◽  
Author(s):  
S Saito ◽  
T Morii ◽  
H Umekage ◽  
K Makita ◽  
K Nishikawa ◽  
...  
Keyword(s):  
T Cells ◽  
Cord Blood ◽  
Nk Cells ◽  
Mononuclear Cells ◽  
Interleukin 2 ◽  
Gamma Chain ◽  
Blood Lymphocytes ◽  
Interleukin 2 Receptor ◽  
Significant Difference ◽  
Cord Blood Lymphocytes

Lymphocytes in umbilical cord blood and neonatal peripheral blood have been shown to have less ability in an immune reaction. In our present experimental approach to address this issue, we made use of the cord blood of full-term birth infants to investigate the expression of the interleukin- 2 receptor gamma (IL-2Rgamma) chain that is shared with receptors for IL-4, IL-7, IL-9, and IL-15 as well as IL-2. The gamma chain expression in cord blood lymphocytes was about one-third that in the lymphocytes of adults, whereas no significant difference between cord blood and adult monocytes was observed. A reduced expression of the gamma chain was observed in all of the CD4+ T cells, CD8+ T cells, gamma-delta T cells, B cells, CD16+ natural killer (NK) cells, and CD56(bright) NK cells of the cord blood lymphocytes. The reduced gamma chain expression reached two-thirds of that in adults after 3 days of culture in vitro and in infants 3 days after birth, thus implying that the increase in the gamma chain may significantly contribute to the prevention of neonatal infection.

scholarly journals TLR agonist combinations that stimulate Th type I polarizing responses from human neonates

2018 ◽  
Vol 24 (4) ◽  
pp. 240-251 ◽  
Author(s):  
Naveen Surendran ◽  
Andrea Simmons ◽  
Michael E Pichichero
Keyword(s):  
Cord Blood ◽  
Lipid A ◽  
Mononuclear Cells ◽  
Poly I:C ◽  
Type I ◽  
Monophosphoryl Lipid A ◽  
Tnf Α ◽  
Tlr Agonist ◽  
Ifn Γ ◽  
Childhood Vaccines

Each year millions of neonates die due to vaccine preventable infectious diseases. Our study seeks to develop novel neonatal vaccines and improve immunogenicity of early childhood vaccines by incorporating TLR agonist-adjuvant combinations that overcome the inherent neonatal Th2 bias and stimulate Th1 polarizing response from neonatal APCs. We systematically stimulated cord blood mononuclear cells with single and multiple combinations of TLR agonists and measured levels of IL-12p70, IFN-γ, IFN-α, IL-10, IL-13, TNF-α, IL-6 and IL-1β from cell culture supernatants. APC-specific surface expression levels of costimulatory markers CD40, CD83 and PD-L1 were assessed by flow cytometry. Whole blood assays were included to account for the effect of plasma inhibitory factors and APC intracellular TNF-α and IL-12p40 secretions were measured. We found robust Th1 polarizing IL-12p70, IFN-γ and IFN-α responses when cord blood APCs were stimulated with TLR agonist combinations that contained Poly I:C, Monophosphoryl Lipid A (MPLA) or R848. Addition of class A CpG oligonucleotide (ODN) to Th1 polarizing TLR agonist combinations significantly reduced cord blood IL-12p70 and IFN-γ levels and addition of a TLR2 agonist induced significantly high Th2 polarizing IL-13. Multi-TLR agonist combinations that included R848 induced lower inhibitory PD-L1 expression on cord blood classical dendritic cells than CpG ODN-containing combinations. Incorporation of combination adjuvants containing TLR3, TLR4 and TLR7/8 agonists to neonatal vaccines may be an effective strategy to overcome neonatal Th2 bias.

C-C chemokine profile of cord blood mononuclear cells: selective defect in RANTES production

Blood ◽  
2000 ◽  
Vol 95 (2) ◽  
pp. 715-718 ◽  
Author(s):  
Deepa Hariharan ◽  
Wenzhe Ho ◽  
Joann Cutilli ◽  
Donald E. Campbell ◽  
Steven D. Douglas
Keyword(s):  
Cord Blood ◽  
Mononuclear Cells ◽  
Flow Cytometric Analysis ◽  
Peripheral Blood Lymphocytes ◽  
Enzyme Linked Immunosorbent Assay ◽  
Blood Lymphocytes ◽  
Perinatal Hiv ◽  
Inflammatory Protein ◽  
Blood Mononuclear Cells ◽  
Cord Blood Mononuclear Cells

Three C-C chemokines inhibit human immunodeficiency virus (HIV) entry into macrophages: macrophage inflammatory protein-1 (MIP-1), MIP-1β, and regulated-upon activation, normal T-cell expressed and secreted (RANTES). We studied the ability of placental cord blood mononuclear cells (CBMC) to secrete these C-C chemokines in comparison to adult blood mononuclear cells (ABMC). CBMC had diminished ability to secrete RANTES, as determined by enzyme-linked immunosorbent assay. Secretion of MIP-1 and MIP-1β were similar in CBMC and ABMC. Whereas MIP-1 and MIP-1β secretion were comparable in monocytes and lymphocytes, RANTES was secreted primarily by lymphocytes. Flow cytometric analysis of RANTES expression showed diminished intracellular RANTES expression in cord blood lymphocytes (CBL) compared to adult (peripheral) blood lymphocytes (ABL). A subset analysis of RANTES-producing CBL and ABL demonstrated that RANTES was produced predominantly by CD8+/CD45RO+ cells. CBL had a reduced proportion of CD8+/CD45RO+ cells compared with ABL, which may account for the diminished RANTES secretion by CBMC. These results may be relevant to the pathogenesis of perinatal HIV infection.

Production of IL-4, IL-2, IFN-γ and TNF-α by peripheral blood lymphocytes in atopic dermatitis

1990 ◽  
Vol 1 (5) ◽  
pp. 389
Author(s):  
Masutaka Furue ◽  
Takenori Takahashi ◽  
Fuyuki Ogata ◽  
Hidemi Nakagawa ◽  
Yasumasa Ishibashi ◽  
...  
Keyword(s):  
Atopic Dermatitis ◽  
Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Blood Lymphocytes ◽  
Tnf Α ◽  
Ifn Γ
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