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J D Tissot

Publications and source records attributed to J D Tissot.

14 recordsLinked to original sources
PubMed1992

Polypeptide marker and disease patterns found while mapping proteins in ascitis.

To assess the protein composition of ascitis, 28 samples of ascitic fluid were obtained from patients admitted to Geneva University Hospital. The samples were analysed randomly and blindly by high-resolution two-dimensional polyacrylamide gel electrophoresis. The final visual evaluation was compared with the discharged summary and diagnosis. The protein pattern of ascitis was, as expected, very similar to normal or diseased plasma, with the exception of two spots which were present in ascitic fluids but not in the 200 plasma samples analyzed in parallel. After microsequencing, they proved to be beta-fibrinogen fragments. Several diseases showed distinct patterns, especially acute pancreatitis. A group of intense spots with an apparent molecular mass between those of alpha 1-antitrypsin and beta-haptoglobin were found in all ascitic fluid from pancreatitis cases (six patients). These spots had isoelectric points similar to those of alpha 1-antitrypsin and beta-haptoglobin and microsequencing revealed that they were three different fragments of alpha 1-antitrypsin.

Acute Disease↗
PubMed1992

Clonality of cold agglutinins in patients with hemolytic anemia: an analysis by high-resolution two-dimensional gel electrophoresis.

High-resolution two-dimensional gel electrophoresis (2-DGE) was used to analyse plasma samples and partially purified cold agglutinins (CA) obtained from two selected patients. Both presented an acute hemolytic anemia with CA of high thermal amplitude, normal immunoglobulin levels, no detectable paraproteinemia, and no clinical evidence of a malignant B-cell disorder. The electrophoretograms of their plasma showed evident alternations of the "normal" protein profile, which were directly related to hemolysis (absence of the spots of haptoglobin and in one case of those of hemopexin), but no monoclonal gammopathy. The electrophoretograms of their purified CA revealed two clearly different spot patterns respectively corresponding to a monoclonal IgM and to polyclonal IgM. These results show that the clonality of CA associated with hemolytic anemia can be easily determined by 2-DGE. This technique may be very useful to discriminate chronic cold agglutinin disease in the early phase from "parainfectious" CA.

Adult↗
PubMed1992

Human liver protein map: a reference database established by microsequencing and gel comparison.

This publication establishes a reference human liver protein map obtained with immobilized pH gradients. By microsequencing, 57 spots or 42 polypeptide chains were identified. By protein map comparison and matching (liver, red blood cell and plasma sample maps), 8 additional proteins were identified. The new polypeptides and previously known proteins are listed in a table and/or labeled on the protein map, thus providing a human liver two-dimensional gel database. This reference map can be used to identify protein spots on other samples such as rectal cancer biopsies.

Amino Acid Sequence↗
PubMed1992

Clonal imbalances of serum immunoglobulins after allogeneic bone marrow transplantation: an analysis by high-resolution two-dimensional gel electrophoresis.

The clonality pattern of immunoglobulins (Igs) produced after allogeneic bone marrow transplantation (BMT) was studied by high-resolution two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) of serum samples and purified Igs. With this technique, the light (L) chain of a monoclonal Ig usually appears as a single spot. Thus, the degree of clonal diversity of the functional B cells can be appreciated by the electrophoretic pattern of the serum L chains. Furthermore, 2D-PAGE allows a semi-quantitative determination of prominent Ig clones according to the size of L chain spots. We found that serum electrophoretograms of 8/19 patients after BMT (5-9 months) revealed L chain patterns which were similar to those of normal polyclonal Igs, that is, less than five distinguishable small spots among a cloud-like indiscrete L chain spots region ('polyclonal' pattern). A spectrum of clonal abnormalities was observed on the electrophoretograms of 11/19 patients: in five patients, multiple small L chain spots (corresponding to Ig concentrations between 0.2 and 2 g/l) were detected ('oligoclonal' pattern), whereas in six others, 'typical' monoclonal Igs (Ig concentrations > 2 g/l) were observed with (3/19 patients) or without (3/19 patients) multiple small clonal components. Sequential analysis of serum obtained from patients at different times after BMT revealed that imbalanced clonal reconstitution was transient and evolved towards apparently normal polyclonal Ig production. Our observations show that the development of clonal 'gammopathies' after BMT is a frequent, but not obligatory phenomenon. It may reflect a transient restriction of the B cell repertoire either through a limited outgrowth of precursor cells or through selective antigenic pressures.

Antibody Diversity↗
PubMed1992

Monoclonal gammopathy in a 30 weeks old premature infant.

The occurrence of monoclonal gammopathy in childhood is extremely rare. This report describes the presence of a monoclonal immunoglobulin in a 30 week old premature infant, incidentally discovered by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) during an ongoing study of the plasma/serum protein development. Comparative analysis of the electrophoretogram of the infant with 'reference' protein maps revealed the presence of an 'abnormal' immunoglobulin light chain spot. A spot having an identical apparent molecular weight and isoelectric point was also detected after 2D-PAGE of the mother's plasma and its Protein-A purified immunoglobulin fraction. The observation of a monoclonal gammopathy in a premature infant, most likely transmitted from his mother, highlights the potential usefulness of 2D-PAGE in the clinical laboratory.

Blood Protein Electrophoresis↗
PubMed1991

[Persistent B-cell polyclonal lymphocytosis: a benign lymphoproliferative syndrome].

A further case of persistent polyclonal B-cell lymphocytosis is reported. This recently identified distinct clinicopathologic entity is frequently associated with the presence of the HLA-DR7 antigen. It follows a benign course and has so far been reported only in women smokers. The disorder is characterized by mild chronic peripheral lymphocytosis, the presence of characteristic binucleate lymphocytes on peripheral blood smears, and a polyclonal increase in serum IgM. In some cases, lymphadenopathies and/or splenomegaly are observed. Surface marker studies of peripheral lymphocytes demonstrate the polyclonal B-cell nature of this entity.

Adult↗
PubMed1991

[Transient episode of erythrocytic autoimmunization in a patient alloimmunized at the end of multiple transfusions].

Delayed hemolytic reaction is a well recognized hazard of blood transfusion and occurs mainly in recipients with alloantibodies, due to sensitization to red cell antigens by previous transfusion or pregnancy. Less frequently, such a reaction may be associated with the presence of red cell autoantibodies appearing after alloimmunization. We report the observation of a patient with chronic myelomonocytic leukemia, who presented a transitory episode of delayed hemolysis after multiple transfusions. At first, anti-Kell alloantibodies were identified and, two weeks later, warm "broad-specific" autoantibodies were detected by direct and indirect antiglobulin tests. The anemia was associated with unstable angina. The patient received multiple transfusions and prednisone. After 3 weeks, the transfusion needs diminished and the autoantibodies progressively disappeared from the serum. The direct antiglobulin test was negative 4 months later. This observation illustrates the poorly understood relationships between erythrocytic alloantigen exposure and red cell autoimmunization.

Aged↗
PubMed1991

High-resolution two-dimensional protein electrophoresis of pathological plasma/serum.

The potential usefulness of an optimalized high-resolution two-dimensional gel electrophoresis (2-DGE) protocol was studied by comparative analysis of plasma/serum obtained from apparently healthy individuals and from patients with a few selected known diseases. Despite their apparent complexity, patient electrophoretograms revealed readily detectable modifications of the 'reference' protein profile for those selected diseases (listed below). Abnormal profiles were characterized by presence or absence of particular spots, by reduction or enlargement of spot size, or by alterations of spot microheterogeneity. Combinations of several modifications enabled different 'disease-associated spot pattern' to be distinguished on the protein maps of patients with: monoclonal gammopathies, hypogammaglobulinemia, hepatic failure, chronic renal failure and hemolytic anemia. This study demonstrates that identification of plasma/serum protein alterations by 2-DGE allows a few selected diseases to be diagnosed solely on the basis of protein map modifications.

Agammaglobulinemia↗
PubMed1989

[Two-dimensional electrophoretic study of monoclonal gammopathies and cold agglutinins associated with a chronic hemolytic anemia].

Two-dimensional gel electrophoresis (2-DGE) associated with a very sensitive silver staining is one of the most powerful technique available for protein analysis. Proteins are separated in the first dimension by isoelectric focusing in a pH gradient, and in the second dimension by polyacrylamide gel electrophoresis. Proteins are thus separated according to their charge and to their size. The high resolution of 2-DGE allows the analysis of complex mixture of proteins with a great reproducibility. More than hundred different proteins can be individualized after 2-DGE of plasma or serum samples. Using this technique, we analysed samples obtained from normal donors and from patients with different gammopathies. Our results confirm that the heavy chains of immunoglobulins are easily detected and that the alpha, gamma and mu chains are clearly differentiated from one another. The typical polyclonal heavy and light chains appear as indiscrete spots after staining of the gels. On the contrary, the monoclonal chains appear as discrete spots. Monoclonal heavy chains have a restricted charge heterogeneity. We also analysed cold agglutinins, partially purified from plasma of three patients showing evidence of chronic hemolysis, a positive direct antiglobulin test of the complement type and anti-I autoantibodies. The 2-DGE have clearly demonstrated that these cold agglutinins were monoclonal IgM, when these later were not detectable by immunofixation electrophoresis. We think that the 2-DGE is a very usefull specialized aid to the analysis of such particular immunohematological problems.

Agglutinins↗
PubMed1988

[Acquired hemophilia. Apropos of a case report with a fatal course].

We report the case of an 86-year-old female with an unremarkable past history who suddenly developed severe and generalized bleeding. Laboratory work-up revealed prolonged aPTT, a factor VIII level of 7% and the presence of a factor VIII inhibitor. In spite of whole blood, factor VIII concentrates and immunoglobulin administration, the patient died of multiple hemorrhages on the third day after admission. At autopsy no causal or associated pathology could be found. Chromatographic separation of plasma immunoglobulins showed that the anti-factor VIII activity was associated with the patient's IgG. This inhibition was partially corrected in vitro by injectable immunoglobulin preparation. The case report stresses the discrepancy between the measured levels of inhibitor and the severity of the bleeding; our in vitro results may shed light on the mode of action of intravenous immunoglobulins.

Aged↗
PubMed1988

Protein heterogeneity of lipoprotein particles containing apolipoprotein A-I without apolipoprotein A-II and apolipoprotein A-I with apolipoprotein A-II isolated from human plasma.

The protein heterogeneity of fractions isolated by immunoaffinity chromatography on anti-apolipoprotein A-I and anti-apolipoprotein A-II affinity columns was analyzed by high resolution two-dimensional gel electrophoresis. The two-dimensional gel electrophoresis profiles of the fractions were analyzed and automatically compared by the computer system MELANIE. Fractions containing apolipoproteins A-I + A-II and only A-I as the major protein components have been isolated from plasma and from high density lipoproteins prepared by ultracentrifugation. Similarities between the profiles of the fractions, as indicated by two-dimensional gel electrophoresis, suggested that those derived from plasma were equivalent to those from high density lipoproteins (HDL), which are particulate in nature. The established apolipoproteins (A-I, A-II, A-IV, C, D, and E) were visible and enriched in fractions from both plasma and HDL. However, plasma-derived fractions showed a much greater degree of protein heterogeneity due largely to enrichment in bands corresponding to six additional proteins. They were present in trace amounts in fractions isolated from HDL and certain of the proteins were visible in two-dimensional gel electrophoresis profiles of the plasma. These proteins are considered to be specifically associated with the immunoaffinity-isolated particles. They have been characterized in terms of Mr and pI. Computer-assisted measurements of protein spot-staining intensities suggest an asymmetric distribution of the proteins (as well as the established apolipoproteins), with four showing greater prominence in particles containing apolipoprotein A-I but no apolipoprotein A-II.

Apolipoprotein A-I↗
PubMedDate not supplied

Plasma protein map: an update by microsequencing.

The reference plasma protein map, obtained with immobilized pH gradients in the first dimension of two-dimensional electrophoresis, is presented. By microsequencing, more than 40 polypeptide chains were identified. The new polypeptides and previously known proteins are listed in a table and labeled on the protein map, thus providing an update of the human plasma two-dimensional gel database.

Amino Acid Sequence↗