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Isolation and Characterization of RNA from Lymphocytes of Chronic Lymphocytic Leukaemia
Author(s) -
Deutsch A.,
Nordé Å.
Publication year - 1971
Publication title -
scandinavian journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.904
H-Index - 84
eISSN - 1600-0609
pISSN - 0036-553X
DOI - 10.1111/j.1600-0609.1971.tb01961.x
Subject(s) - phytohaemagglutinin , rna , microbiology and biotechnology , ribosomal rna , biology , differential centrifugation , polynucleotide , agarose , lymphocyte , biochemistry , immunology , gene
RNA was isolated from lymphocytes of the peripheral blood of patients with chronic lymphocytic leukaemia under conditions previously shown (Cline 1966a) to result in undegraded RNA. At the same time, responsiveness of the lymphocytes to PHA was measured by determination of the incorporation of 3 H‐thymidine in culture. The isolated RNA was characterized by sucrose density gradient centrifugation, gel chromatography on 2% agarose, electrophoresis on mixed polyacrylamide‐agarose gels, and fractionation with 3 M NaCl. RNA isolated from leukaemic lymphocytes unresponsive to phytohaemagglutinin showed a pattern identical to that obtained from normal lymphocytes and liver. RNA preparations from leukaemic lymphocytes responsive to phytohaemagglutinin gave a diverging pattern, characterized by the presence of large amounts of relatively low molecular or polydisperse RNA. Chronic lymphocytic leukaemia lymphocytes not responsive to phytohaemagglutinin gave, in analogy to normal lymphocytes and liver, 28s + 18s rRNA in amounts corresponding to 75 to 85% of total extractable RNA, whereas leukaemic lymphocytes responsive to phytohaemagglutinin yielded only 40–60% of the total extractable RNA in the form of 28s + 18s RNA.