Naturally Occurring Human Antibody to Neuraminidase-Treated Human Lymphocytes. Antibody Levels in Normal Subjects, Cancer Patients, and Subjects With Immunodeficiency

Lymphocytes from normal subjects and patients with B-chronic lymphocytic leukemia (B-CLL) show functional responses to extracellular ATP characteristic of the P2X7receptor (previously termed P2Z). These responses include opening of a cation-selective channel/pore that allows entry of the fluorescent dye ethidium and activation of a membrane metalloprotease that sheds the adhesion molecule L-selectin. The surface expression of P2X7receptors was measured in normal leucocytes, platelets, and B-CLL lymphocytes and correlated with their functional responses. Monocytes showed four- to fivefold greater expression of P2X7than B, T, and NK lymphocytes, whereas P2X7expression on neutrophils and platelets was weak. All cell types demonstrated abundant intracellular expression of this receptor. All 12 subjects with B-CLL expressed lymphocyte P2X7at about the same level as B lymphocytes from normal subjects. P2X7function, measured by ATP-induced uptake of ethidium, correlated closely with surface expression of this receptor in normal and B-CLL lymphocytes and monocytes ( n = 47, r = 0.70; P< 0.0001). However, in three patients the ATP-induced uptake of ethidium into the malignant B lymphocytes was low or absent. The lack of P2X7function in these B lymphocytes was confirmed by the failure of ATP to induce Ba2+uptake into their lymphocytes. This lack of function of the P2X7receptor resulted in a failure of ATP-induced shedding of L-selectin, an adhesion molecule that directs the recirculation of lymphocytes from blood into the lymph node.

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Survival in Early Breast Cancer Patients Is Favorably Influenced by a Natural Humoral Immune Response to Polymorphic Epithelial Mucin

Journal of Clinical Oncology ◽

10.1200/jco.2000.18.3.574 ◽

2000 ◽

Vol 18 (3) ◽

pp. 574-574 ◽

Cited By ~ 153

Author(s):

S. von Mensdorff-Pouilly ◽

A.A. Verstraeten ◽

P. Kenemans ◽

F.G. M. Snijdewint ◽

A. Kok ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Patients ◽

Early Breast Cancer ◽

Humoral Response ◽

Disease Specific Survival ◽

Breast Cancer Patients ◽

Antibody Levels ◽

Pretreatment Serum ◽

Test Result ◽

Disease Specific

PURPOSE: Polymorphic epithelial mucin (PEM or MUC1) is being studied as a vaccine substrate for the immunotherapy of patients with adenocarcinoma. The present study analyzes the incidence of naturally occurring MUC1 antibodies in early breast cancer patients and relates the presence of these antibodies in pretreatment serum to outcome of disease.MATERIALS AND METHODS: We measured immunoglobulin G (IgG) and immunoglobulin M (IgM) antibodies to MUC1 with an enzyme-linked immunoassay (PEM.CIg), which uses a MUC1 triple-tandem repeat peptide conjugated to bovine serum albumin, in pretreatment serum samples obtained from 154 breast cancer patients (52 with stage I disease and 102 with stage II) and 302 controls. The median disease-specific survival time of breast cancer patients was 74 months (range, 15 to 118 months). A positive test result was defined as MUC1 IgG or IgM antibody levels equal to or greater than the corresponding rounded-up median results obtained in the total breast cancer population.RESULTS: A positive test result for both MUC1 IgG and IgM antibodies in pretreatment serum was associated with a significant benefit in disease-specific survival in stage I and II (P = .0116) breast cancer patients. Positive IgG and IgM MUC1 antibody levels had significant additional prognostic value to stage (P = .0437) in multivariate analysis. Disease-free survival probability did not differ significantly. However, stage II patients who tested positive for MUC1 IgG and IgM antibody and who relapsed had predominantly local recurrences or contralateral disease, as opposed to recurrences at distant sites in the patients with a negative humoral response (P = .026).CONCLUSION: Early breast cancer patients with a natural humoral response to MUC1 have a higher probability of freedom from distant failure and a better disease-specific survival. MUC1 antibodies may control hematogenic tumor dissemination and outgrowth by aiding the destruction of circulating or seeded MUC1-expressing tumor cells. Vaccination of breast cancer patients with MUC1-derived (glyco)peptides in an adjuvant setting may favorably influence the outcome of disease.

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Generation of primary antigen-specific human cytotoxic T lymphocytes in human/mouse radiation chimera

Blood ◽

10.1182/blood.v88.2.721.bloodjournal882721 ◽

1996 ◽

Vol 88 (2) ◽

pp. 721-730 ◽

Cited By ~ 30

Author(s):

H Segall ◽

I Lubin ◽

H Marcus ◽

A Canaan ◽

Y Reisner

Keyword(s):

T Cells ◽

T Cell ◽

Human Lymphocytes ◽

Scid Mice ◽

Human Antibody ◽

Adoptive Therapy ◽

Activation Markers ◽

Human T Cell ◽

Human T Cells

Severe combined immunodeficient (SCID) mice are increasingly used as hosts for the adoptive transfer of human lymphocytes. Human antibody responses can be obtained in these xenogeneic chimeras, but information about the functionality of the human T cells in SCID mice is limited and controversial. Studies using human peripheral blood lymphocytes (PBL) injected intraperitoneally (IP) into SCID mice (hu-PBL-SCID mice) have shown that human T cells from these chimeras are anergic and have a defective signaling via the T-cell receptor. In addition, their antigenic repertoire is limited to xenoreactive clones. In the present study, we tested the functionality of human T cell in a recently described chimeric model. In this system, BALB/c mice are conditioned by irradiation and then transplanted with SCID bone marrow, followed by IP injection of human PBL. Our experiments demonstrated that human T cells, recovered from these hu-PBL-BALB mice within 1 month posttransplant, proliferated and expressed activation markers upon stimulation with anti-CD3 monoclonal antibody. A vigorous antiallogeneic human cytotoxic T-lymphocyte (CTL) response could be generated in these mice by immunizing them with irradiated allogeneic cells. Moreover, anti-human immunodeficiency virus type 1 (HIV-1) Net- specific human CTLs could be generated in vivo from naive lymphocytes by immunization of mouse-human chimeras with a recombinant vaccinia-nef virus. This model may be used to evaluate potential immunomodulatory drugs or cytokines, and could provide a relevant model for testing HIV vaccines, for production of antiviral T-cell clones for adoptive therapy, and for studying human T-cell responses in vivo.

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Plasma lipid-bound sialic acid and carcinoembryonic antigen in cancer patients.

Clinical Chemistry ◽

10.1093/clinchem/27.10.1737 ◽

1981 ◽

Vol 27 (10) ◽

pp. 1737-1739 ◽

Cited By ~ 22

Author(s):

A M Dnistrian ◽

M K Schwartz

Keyword(s):

Sialic Acid ◽

Carcinoembryonic Antigen ◽

Cancer Patients ◽

Lipid Extraction ◽

Plasma Lipid ◽

Normal Subjects ◽

Breast Cancer Patients ◽

Lung Cancer Patients ◽

Total Lipid Extraction ◽

The Individual

Abstract We evaluated lipid-bound sialic acid as a "marker" in cancer patients and assessed the individual and combined value of lipid-bound sialic acid and carcinoembryonic antigen determinations in these patients. Plasma was sampled from 62 normal subjects and 125 cancer patients. Lipid-bound sialic acid was determined by the resorcinol method after total lipid extraction and isolation of the sialolipid fraction from plasma. Neither marker was increased in many breast cancer patients. Carcinoembryonic antigen was increased more commonly and to a greater degree in colon cancer patients and seems to be the preferred marker. Both markers were increased in lung cancer patients and their combined evaluation improved the rate of detection. Lipid-bound sialic acid was increased in more patients with leukemias, lymphomas, Hodgkin's disease, and melanomas, suggesting that it may be a useful biochemical marker in these types of cancer.

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The Separation and 51Chromium Labeling of Human Lymphocytes With In Vivo Studies of Survival and Migration

Blood ◽

10.1182/blood.v38.3.360.360 ◽

1971 ◽

Vol 38 (3) ◽

pp. 360-371 ◽

Cited By ~ 30

Author(s):

PETER HERSEY

Keyword(s):

Human Lymphocytes ◽

Normal Subjects ◽

The Body ◽

In Vivo Studies ◽

Human Beings ◽

Lymphatic Leukemia ◽

Circulating Lymphocytes ◽

Relative Inability ◽

And Migration

Abstract This study looks at the application of 51Cr labeling of lymphocytes as a method of obtaining in vivo information about the lymphocyte in human beings. Lymphocytes were separated from whole blood by methods based on isopycnic and rate zonal centrifugation techniques and the conditions for 51Cr uptake by the separated lymphocytes standardized to enable a known amount of radioactivity to be injected into the subjects under study. The uptake of the label into various sites in the body was studied by the means of surface probes linked synchronously to a digital printout device and the survival in the circulation estimated by scintillation counting of blood samples taken at various times after injection of the label. The in vivo studies of survival and migration in 10 normal subjects show an initial rapid clearance of cells from the circulation associated with an uptake of cells into spleen and liver sites, and to a lesser extent, into sites over bone marrow and the abdomen. Survival of the circulating lymphocytes after this period appears to be relatively short, with a half-life of 1.7 days. As the available evidence suggests, this short life may be due to the differential trapping of short-lived lymphocytes in the circulation at the expense of the long-lived lymphocytes. Kinetic interpretations of the data indicate an inverse exponential uptake of cells into the sites studied, and the decline over the organs appears to follow the death rate of the cells in the body as a whole. Comparisons with studies in patients having chronic lymphatic leukemia show a relative inability of leukemic lymphocytes to leave the circulation and enter some sites in the body. These preliminary studies indicate the potential of 51Cr labeling as a useful clinical research tool in the study of lymphocytes in human beings.

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Comparison of simple acid-ethanol precipitation with gel exclusion chromatography for measuring leptin binding in serum of normal subjects and cancer patients

Annals of Clinical Biochemistry International Journal of Laboratory Medicine ◽

10.1258/000456303763046157 ◽

2003 ◽

Vol 40 (2) ◽

pp. 185-187 ◽

Cited By ~ 2

Author(s):

Nigel B. Jamieson ◽

Donald C. McMillan ◽

Duncan J. F. Brown ◽

A. Michael Wallace

Keyword(s):

Cancer Patients ◽

Normal Subjects ◽

Free Form ◽

Careful Evaluation ◽

Serum Samples ◽

Ethanol Precipitation ◽

Significant Difference ◽

Exclusion Chromatography ◽

Analysis Of Results ◽

Assay Precision

Background: In humans, leptin circulates in a free form and is also bound to macromolecules. The aim of the present study was to compare a rapid acid-ethanol precipitation (AEP) method of measuring bound leptin with the more laborious gel exclusion chromatography (GEC) reference procedure. Serum samples collected from healthy subjects and cancer patients were used in this comparison. Methods: AEP and GEC methods for measuring leptin binding in serum (from 14 healthy volunteers and 14 patients with advanced non-small cell lung cancer) were adapted from previously published procedures. Results: Intra- and inter-assay precision of the AEP method were 6% ( n = 10) and 8% ( n = 10), respectively. Bland-Altman analysis of results obtained from the AEP and GEC methods indicated no significant difference in healthy controls. However, significantly higher results were obtained by the AEP method in the cancer patients. Conclusions: Evaluation of the AEP method revealed that on examination of normal subjects the method was less precise than had previously been reported. Moreover, the method gave differing results in the cancer patients when compared with the GEC method. This study indicates that careful evaluation of any new method for measuring leptin binding requires comparison with a GEC method using the sample matrix of interest.

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Concentration of glucocorticoid receptor sites in normal human lymphocytes.

Clinical Chemistry ◽

10.1093/clinchem/32.1.80 ◽

1986 ◽

Vol 32 (1) ◽

pp. 80-83 ◽

Cited By ~ 4

Author(s):

A E Steiner ◽

J L Wittliff

Keyword(s):

Glucocorticoid Receptor ◽

Human Lymphocytes ◽

Normal Subjects ◽

Mean Value ◽

Whole Cell ◽

Receptor Sites ◽

Human T Cells ◽

Cell Assays ◽

Normal Human ◽

The Mean

Abstract We used a whole-cell glucocorticoid receptor assay to examine characteristics of the glucocorticoid receptor in the lymphocytes of normal human donors. We measured binding of [3H]dexamethasone to the lymphocytes of four different donors on several different occasions; the variation about the mean for the assays was +/- 15%. Whole-cell assays in 15 normal subjects showed a mean value of 6.18 fmol/10(6) cells or 3722 sites per cell, with a somewhat higher level in men (7.67 fmol/10(6) cells, or 4620 sites per cell) than women (4.48 fmol/10(6) cells, or 2698 sites per cell). We saw no correlation between donor age and receptor values, in either group. Assays in which we used [3H]prednisolone demonstrated similar binding properties as with [3H]dexamethasone. The mean glucocorticoid receptor value for normal human T-cells from three donors was 2.52 fmol/10(6) cells, or 1518 sites per cell.

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Urinary Sulfur Partition in Normal Men and in Cancer Patients

Clinical Chemistry ◽

10.1093/clinchem/3.4.257 ◽

1957 ◽

Vol 3 (4) ◽

pp. 257-262 ◽

Cited By ~ 7

Author(s):

Daphne B Papadopoulou

Keyword(s):

Cancer Patients ◽

Normal Subjects ◽

The Body ◽

Normal Men ◽

Total Sulfate

Abstract 1. The partition of inorganic and total sulfate and total and neutral sulfur has been estimated in 10 normal men and 27 cancer patients. 2. The excretion of inorganic and total sulfate was found to be decreased and the neutral sulfur fraction increased in cancer patients more than in the normal subjects. 3. From the data above, it is suggested that there may be a decrease in the rate of the intracellular oxidation substances in the body of cancer patients.

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