Studies on the Immunology and Laboratory Investigation of B Cell Neoplasia

Sinclair, David (1986) Studies on the Immunology and Laboratory Investigation of B Cell Neoplasia. PhD thesis, University of Glasgow.

Abstract

A scheme for the detection and identification of serum and urinary monoclonal immunoglobulins using agarose isoelectric focusing is described. Conditions required for satisfactory isoelectric focusing of all classes of immunoglobulin are shown. Comparisons in sensitivity between isoelectric focusing (IEF) and immuno-isoelectric focusing (IIEF) with other techniques commonly used to detect and identify paraproteins showed that the two techniques were 10-40 times more sensitive compared with routine immunoelectrophoresis. The use of IIEF in a number of clinical situations is described. On occasion, patients respond to therapy for myeloma with complete disappearance of paraprotein from serum as judged by immunoelectrophoresis. The use of IIEF to these 'remission' patients shows that in a group of 27 such patients, 16 had paraproteinaemia detectable by IIEF compared with only 7 by zonal electrophoresis followed by immunofixation, another technique commonly used in this regard. The first demonstration of 'in vivo' paraproteinaemia in a case diagnosed as 'non-secretcry' myeloma is described. The concentration of the paraprotein is given along with treatment dates showing a steady paraprotein concentration before the the patients death. The use of IIEF for the detection of paraproteinaemia is described in two cases of myelomatosis involving intact immunoglobulin molecules; one case involving Bence-Jones myeloma and one case involving a monoclonal gammopathy of undetermined significance, along with treatment date where appropriate, thus charting the progress of the diseases. The incidence and possible significance of serum Bence-Jones protein was investigated. The detectable incidence of Bence- Jones protein in the sera of myeloma patients was found to be equivalent to the incidence in concentrated urine when tested by IIEF. In a series of 25 myeloma patients, IIEF showed the incidence of Bence-Jones proteinaernia to be 68% compared with a detectable incidence of Bence-Jones proteinuria of 32-64% by immunoelectrophoresis and IIEF depending on the concentration of the urine. These results suggest that in view of the variability of factors governing urinary Bence-Jones protein levels, monitoring of serum Bence-Jones protein by IIEF should be carried out in conjunction with measurements of urinary Bence-Jones protein in myeloma patients. The detection of a paraprotein, previously unsuspected using routine techniques, in several cases of solitary plasmacytoma is described. The use of IIEF in cases such as these may offer an additional means of assessing disease progress and response to chemotherapy. A method for quantifying paraproteins in serum and urine using scanning densitometric analysis of IEF and IIEF tracks is described, thus enabling paraprotein concentrations to be monitored from low levels up to and including those quantifiable using conventional techniques. Immuno-isoelectric focusing was also used to screen a total of 56 patients with chronic lymphocytic leukaemia for the presence of serum paraproteinaemia. A total of 34 (61%) of patients had paraproteins. These were mostly of IgM class, but paraproteins of other isotypes were detected. The serum paraprotein isotypes were compared with the isotypes of cytoplasmic immunoglobulin of peripheral blood lymphocytes from the same patients. A strong correlation emerged between them suggesting that the paraproteins originated from the neoplastic clone. Density gradient ultracentrifugation of 8 sera from CLL patients with IgM paraproteinaemia showed that for the most part the IgM paraproteins were 1 9S in size, but one patient was found to have both 19S and 8S monoclonal IgM. The cellular origin of the paraproteins (monoclonal IgM lambda & IgD lambda ) in the serum of one patient was investigated using an anti-idiotype antibody raised against the IgM paraprotein. This work showed that both paraproteins shared a common idiotype, i. e. both were secreted from the same clone of B cells. These results were used to support the argument that there is an incomplete maturation defect in chronic lymphocytic leukaemia leading to a limited secretory capacity in the majority of cases. In addition, a total of 200 individuals without history of B cell neoplasm were tested for the presence of abnormal immunoglobulin profiles. A total of 22 people had monoclonal paraproteins with 24 people having IIEF traces which corresponded to cligoclonal immunoglobulins. The relevance of these results is discussed. The presence of serum monoclonal immunoglobulin in non-Hodgkin's lymphoma and also in Hodgkins disease is demonstrated by IIEF. The significance of paraproteinaemia in the latter disease is discussed.

• Item Type: Thesis (PhD)
• Qualification Level: Doctoral
• Keywords: Immunology
• Date of Award: 1986
• Depositing User: Enlighten Team
• Unique ID: glathesis:1986-77330
• Copyright: Copyright of this thesis is held by the author.
• Date Deposited: 14 Jan 2020 11:53
• Last Modified: 14 Jan 2020 11:53
• URI: https://theses.gla.ac.uk/id/eprint/77330