Flow Cytometric Measurement of Intracellular Cytokines Detects Immune Responses in MUC1 Immunotherapy
The detection of tumor-specific T cells in immunized cancer patients usually relies on lengthy and difficult CTL assays; we now report on flow cytometry to detect the intracellular cytokines interleukin 2 (IL-2), IL-4, IFN-γ, and tumor necrosis factor α (TNF-α) produced by CD4 + CD69 + and CD8 + CD6...
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| Veröffentlicht in: | Clinical cancer research 2000-03, Vol.6 (3), p.829-837 |
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| creator | Karanikas, V Lodding, J Maino, V C McKenzie, I F |
| description | The
detection of tumor-specific T cells in immunized cancer patients
usually relies on lengthy and difficult CTL assays; we now report on
flow cytometry to detect the intracellular cytokines interleukin 2
(IL-2), IL-4, IFN-γ, and tumor necrosis factor α (TNF-α) produced
by CD4 + CD69 + and
CD8 + CD69 + activated T cells after MUC1 antigen
stimulation. Peripheral blood mononuclear cells were obtained
from 12 patients with adenocarcinoma injected with mannan-MUC1;
cells were exposed in vitro for 18 h to MUC1
peptide in the presence of CD28 monoclonal antibody and
Brefeldin; permeabilized cells were used for the expression of
cytokines. After stimulation in vitro with MUC1-variable
number of tandem repeats peptides, CD8 + CD69 + T
cells from all immunized patients generated 3–9 times higher levels of
TNF-α ( P < 0.038) and IFN-γ
( P < 0.010) than did cells from 12 normal
subjects; minor increases in IL-4 occurred. By contrast,
CD4 + CD69 + cells showed no overall alteration in
TNF-α and IFN-γ cytokine production, although in some patients,
their measurement was informative; the measurement of IL-2 was not
useful in either CD4 + CD69 + or
CD8 + CD69 + cells. We conclude that in
MUC1-immunized patients, the measurement of TNF-α and IFN-γ in
activated CD69 + CD8 + T cells may be indicative
of their immune status. |
| format | Article |
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detection of tumor-specific T cells in immunized cancer patients
usually relies on lengthy and difficult CTL assays; we now report on
flow cytometry to detect the intracellular cytokines interleukin 2
(IL-2), IL-4, IFN-γ, and tumor necrosis factor α (TNF-α) produced
by CD4 + CD69 + and
CD8 + CD69 + activated T cells after MUC1 antigen
stimulation. Peripheral blood mononuclear cells were obtained
from 12 patients with adenocarcinoma injected with mannan-MUC1;
cells were exposed in vitro for 18 h to MUC1
peptide in the presence of CD28 monoclonal antibody and
Brefeldin; permeabilized cells were used for the expression of
cytokines. After stimulation in vitro with MUC1-variable
number of tandem repeats peptides, CD8 + CD69 + T
cells from all immunized patients generated 3–9 times higher levels of
TNF-α ( P < 0.038) and IFN-γ
( P < 0.010) than did cells from 12 normal
subjects; minor increases in IL-4 occurred. By contrast,
CD4 + CD69 + cells showed no overall alteration in
TNF-α and IFN-γ cytokine production, although in some patients,
their measurement was informative; the measurement of IL-2 was not
useful in either CD4 + CD69 + or
CD8 + CD69 + cells. We conclude that in
MUC1-immunized patients, the measurement of TNF-α and IFN-γ in
activated CD69 + CD8 + T cells may be indicative
of their immune status.</description><identifier>ISSN: 1078-0432</identifier><identifier>EISSN: 1557-3265</identifier><identifier>PMID: 10741704</identifier><language>eng</language><publisher>United States: American Association for Cancer Research</publisher><subject>Adenocarcinoma - therapy ; Cytokines - metabolism ; Flow Cytometry ; Humans ; Immunization ; Immunotherapy ; Influenza Vaccines - pharmacology ; Interferon-gamma - metabolism ; Interleukin-2 - metabolism ; Leukocytes, Mononuclear - drug effects ; Leukocytes, Mononuclear - immunology ; Leukocytes, Mononuclear - metabolism ; Mannans - immunology ; Mucin-1 - genetics ; Mucin-1 - immunology ; Phytohemagglutinins - pharmacology ; Recombinant Fusion Proteins - immunology ; T-Lymphocytes - drug effects ; T-Lymphocytes - immunology ; T-Lymphocytes - metabolism ; Tetanus Toxoid - pharmacology ; Tumor Necrosis Factor-alpha - metabolism</subject><ispartof>Clinical cancer research, 2000-03, Vol.6 (3), p.829-837</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10741704$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Karanikas, V</creatorcontrib><creatorcontrib>Lodding, J</creatorcontrib><creatorcontrib>Maino, V C</creatorcontrib><creatorcontrib>McKenzie, I F</creatorcontrib><title>Flow Cytometric Measurement of Intracellular Cytokines Detects Immune Responses in MUC1 Immunotherapy</title><title>Clinical cancer research</title><addtitle>Clin Cancer Res</addtitle><description>The
detection of tumor-specific T cells in immunized cancer patients
usually relies on lengthy and difficult CTL assays; we now report on
flow cytometry to detect the intracellular cytokines interleukin 2
(IL-2), IL-4, IFN-γ, and tumor necrosis factor α (TNF-α) produced
by CD4 + CD69 + and
CD8 + CD69 + activated T cells after MUC1 antigen
stimulation. Peripheral blood mononuclear cells were obtained
from 12 patients with adenocarcinoma injected with mannan-MUC1;
cells were exposed in vitro for 18 h to MUC1
peptide in the presence of CD28 monoclonal antibody and
Brefeldin; permeabilized cells were used for the expression of
cytokines. After stimulation in vitro with MUC1-variable
number of tandem repeats peptides, CD8 + CD69 + T
cells from all immunized patients generated 3–9 times higher levels of
TNF-α ( P < 0.038) and IFN-γ
( P < 0.010) than did cells from 12 normal
subjects; minor increases in IL-4 occurred. By contrast,
CD4 + CD69 + cells showed no overall alteration in
TNF-α and IFN-γ cytokine production, although in some patients,
their measurement was informative; the measurement of IL-2 was not
useful in either CD4 + CD69 + or
CD8 + CD69 + cells. We conclude that in
MUC1-immunized patients, the measurement of TNF-α and IFN-γ in
activated CD69 + CD8 + T cells may be indicative
of their immune status.</description><subject>Adenocarcinoma - therapy</subject><subject>Cytokines - metabolism</subject><subject>Flow Cytometry</subject><subject>Humans</subject><subject>Immunization</subject><subject>Immunotherapy</subject><subject>Influenza Vaccines - pharmacology</subject><subject>Interferon-gamma - metabolism</subject><subject>Interleukin-2 - metabolism</subject><subject>Leukocytes, Mononuclear - drug effects</subject><subject>Leukocytes, Mononuclear - immunology</subject><subject>Leukocytes, Mononuclear - metabolism</subject><subject>Mannans - immunology</subject><subject>Mucin-1 - genetics</subject><subject>Mucin-1 - immunology</subject><subject>Phytohemagglutinins - pharmacology</subject><subject>Recombinant Fusion Proteins - immunology</subject><subject>T-Lymphocytes - drug effects</subject><subject>T-Lymphocytes - immunology</subject><subject>T-Lymphocytes - metabolism</subject><subject>Tetanus Toxoid - pharmacology</subject><subject>Tumor Necrosis Factor-alpha - metabolism</subject><issn>1078-0432</issn><issn>1557-3265</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkE1Lw0AURYMoVqt_QWblLvDmI8lkKdFqoUUQux4m6YuJJjNxZkLpvze1FVfvwjlcLu8suqJJksWcpcn5lCGTMQjOZtG1958AVFAQl9FsAoJmIK4iXHR2R4p9sD0G11ZkjdqPDns0gdiaLE1wusKuGzvtfr2v1qAnjxiwCp4s-340SN7QD9b4CbSGrDcFPQIbGnR62N9EF7XuPN6e7jzaLJ7ei5d49fq8LB5WccO4DDGXFMqUCtCH0QhC5pkGnaZAtzVlEngp8oxpKBESySCtsWZSpprnMsmk4PPo_tg7OPs9og-qb_1hvTZoR68yChRymk_i3Ukcyx63anBtr91e_T3mv6lpP5pd61BV2lToHHrUrmpUqriSLOc_nbts-A</recordid><startdate>20000301</startdate><enddate>20000301</enddate><creator>Karanikas, V</creator><creator>Lodding, J</creator><creator>Maino, V C</creator><creator>McKenzie, I F</creator><general>American Association for Cancer Research</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>20000301</creationdate><title>Flow Cytometric Measurement of Intracellular Cytokines Detects Immune Responses in MUC1 Immunotherapy</title><author>Karanikas, V ; Lodding, J ; Maino, V C ; McKenzie, I F</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-h238t-3810b6140a0432e04897a0a6601df12803b4972a0be058206fef2886a39857843</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Adenocarcinoma - therapy</topic><topic>Cytokines - metabolism</topic><topic>Flow Cytometry</topic><topic>Humans</topic><topic>Immunization</topic><topic>Immunotherapy</topic><topic>Influenza Vaccines - pharmacology</topic><topic>Interferon-gamma - metabolism</topic><topic>Interleukin-2 - metabolism</topic><topic>Leukocytes, Mononuclear - drug effects</topic><topic>Leukocytes, Mononuclear - immunology</topic><topic>Leukocytes, Mononuclear - metabolism</topic><topic>Mannans - immunology</topic><topic>Mucin-1 - genetics</topic><topic>Mucin-1 - immunology</topic><topic>Phytohemagglutinins - pharmacology</topic><topic>Recombinant Fusion Proteins - immunology</topic><topic>T-Lymphocytes - drug effects</topic><topic>T-Lymphocytes - immunology</topic><topic>T-Lymphocytes - metabolism</topic><topic>Tetanus Toxoid - pharmacology</topic><topic>Tumor Necrosis Factor-alpha - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Karanikas, V</creatorcontrib><creatorcontrib>Lodding, J</creatorcontrib><creatorcontrib>Maino, V C</creatorcontrib><creatorcontrib>McKenzie, I F</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>Clinical cancer research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Karanikas, V</au><au>Lodding, J</au><au>Maino, V C</au><au>McKenzie, I F</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Flow Cytometric Measurement of Intracellular Cytokines Detects Immune Responses in MUC1 Immunotherapy</atitle><jtitle>Clinical cancer research</jtitle><addtitle>Clin Cancer Res</addtitle><date>2000-03-01</date><risdate>2000</risdate><volume>6</volume><issue>3</issue><spage>829</spage><epage>837</epage><pages>829-837</pages><issn>1078-0432</issn><eissn>1557-3265</eissn><abstract>The
detection of tumor-specific T cells in immunized cancer patients
usually relies on lengthy and difficult CTL assays; we now report on
flow cytometry to detect the intracellular cytokines interleukin 2
(IL-2), IL-4, IFN-γ, and tumor necrosis factor α (TNF-α) produced
by CD4 + CD69 + and
CD8 + CD69 + activated T cells after MUC1 antigen
stimulation. Peripheral blood mononuclear cells were obtained
from 12 patients with adenocarcinoma injected with mannan-MUC1;
cells were exposed in vitro for 18 h to MUC1
peptide in the presence of CD28 monoclonal antibody and
Brefeldin; permeabilized cells were used for the expression of
cytokines. After stimulation in vitro with MUC1-variable
number of tandem repeats peptides, CD8 + CD69 + T
cells from all immunized patients generated 3–9 times higher levels of
TNF-α ( P < 0.038) and IFN-γ
( P < 0.010) than did cells from 12 normal
subjects; minor increases in IL-4 occurred. By contrast,
CD4 + CD69 + cells showed no overall alteration in
TNF-α and IFN-γ cytokine production, although in some patients,
their measurement was informative; the measurement of IL-2 was not
useful in either CD4 + CD69 + or
CD8 + CD69 + cells. We conclude that in
MUC1-immunized patients, the measurement of TNF-α and IFN-γ in
activated CD69 + CD8 + T cells may be indicative
of their immune status.</abstract><cop>United States</cop><pub>American Association for Cancer Research</pub><pmid>10741704</pmid><tpages>9</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1078-0432 |
| ispartof | Clinical cancer research, 2000-03, Vol.6 (3), p.829-837 |
| issn | 1078-0432 1557-3265 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_71010919 |
| source | MEDLINE; American Association for Cancer Research; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Adenocarcinoma - therapy Cytokines - metabolism Flow Cytometry Humans Immunization Immunotherapy Influenza Vaccines - pharmacology Interferon-gamma - metabolism Interleukin-2 - metabolism Leukocytes, Mononuclear - drug effects Leukocytes, Mononuclear - immunology Leukocytes, Mononuclear - metabolism Mannans - immunology Mucin-1 - genetics Mucin-1 - immunology Phytohemagglutinins - pharmacology Recombinant Fusion Proteins - immunology T-Lymphocytes - drug effects T-Lymphocytes - immunology T-Lymphocytes - metabolism Tetanus Toxoid - pharmacology Tumor Necrosis Factor-alpha - metabolism |
| title | Flow Cytometric Measurement of Intracellular Cytokines Detects Immune Responses in MUC1 Immunotherapy |
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