Analysis of immunoglobulin and T cell receptor genes. Part I: Basic and technical aspects

Jj, van Dongen; Il, Wolvers-Tettero

doi:10.1016/0009-8981(91)90246-9

Cited by 207 publications

(175 citation statements)

References 390 publications

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“…26 Control DNA was obtained from cell samples with germline Ig genes. Fifteen micrograms of DNA were digested with the appropriate restriction enzymes (Pharmacia), sizeseparated in 0.7% agarose gels and transferred onto Nytran-13N membranes (Schleicher and Schuell, Dassel, Germany), which were hybridized with 32 P-random oligonucleotide labeled DNA probes as described previously.…”

Section: Southern Blot Analysis Of Ig Genesmentioning

confidence: 99%

“…Fifteen micrograms of DNA were digested with the appropriate restriction enzymes (Pharmacia), sizeseparated in 0.7% agarose gels and transferred onto Nytran-13N membranes (Schleicher and Schuell, Dassel, Germany), which were hybridized with 32 P-random oligonucleotide labeled DNA probes as described previously. 26 Ig heavy chain (IgH) gene rearrangements were detected with the IGHJ6 probe in BglII and combined BamHI/HindIII digests. 26,27 Ig gene rearrangements and deletions were detected with the IGKJ5, IGKC, and IGKDE probes in BglII and combined BamHI/HindIII digests.…”

Section: Southern Blot Analysis Of Ig Genesmentioning

confidence: 99%

“…26 Ig heavy chain (IgH) gene rearrangements were detected with the IGHJ6 probe in BglII and combined BamHI/HindIII digests. 26,27 Ig gene rearrangements and deletions were detected with the IGKJ5, IGKC, and IGKDE probes in BglII and combined BamHI/HindIII digests. 26,28 The IGKDE probe in combination with the IGKC and IGKJ5 probes allows the precise detection of the two types of Ig deletions, ie C deletion and J -C deletion.…”

Section: Southern Blot Analysis Of Ig Genesmentioning

confidence: 99%

“…26,27 Ig gene rearrangements and deletions were detected with the IGKJ5, IGKC, and IGKDE probes in BglII and combined BamHI/HindIII digests. 26,28 The IGKDE probe in combination with the IGKC and IGKJ5 probes allows the precise detection of the two types of Ig deletions, ie C deletion and J -C deletion. 28 Ig gene rearrangements were studied by use of the IGLC3 probe in EcoRI and combined EcoRI/HindIII digests; these probe/enzyme combinations allow the detection of virtually all (Ͼ95%) Ig gene rearrangements.…”

Section: Southern Blot Analysis Of Ig Genesmentioning

confidence: 99%

“…28 Ig gene rearrangements were studied by use of the IGLC3 probe in EcoRI and combined EcoRI/HindIII digests; these probe/enzyme combinations allow the detection of virtually all (Ͼ95%) Ig gene rearrangements. 26,29 Cytogenetic studies Peripheral blood cells were cultured for 4 and 5 days using various mitogens, ie TPA (0.15 g/ml), LPS (100 g/ml) and PHA (0.15 mg/ml). Metaphase arrest using colcemid (0.2 g/ml) and harvesting procedures were performed according to standard protocols.…”

Section: Southern Blot Analysis Of Ig Genesmentioning

confidence: 99%

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Four aged siblings with B cell chronic lymphocytic leukemia

F¹,

Dinkelaar

Hagemeijer

et al. 1997

Leukemia

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All four aged siblings (Ͼ80 years) of one family presented with B cell chronic lymphocytic leukemia (B-CLL). In an attempt to find common characteristics in the four patients, we performed detailed immunological marker analysis, Southern blot analysis of immunoglobulin (Ig) genes, and cytogenetic studies. In three patients clonality of the B-cells could be proven by single Ig light chain expression, but in the fourth patient no Ig light chain expression was detected and clonality of the B cells could only be demonstrated by Southern blot analysis of the Ig genes. Interestingly, in two patients, the Ig gene rearrangement patterns were compatible with the presence of two independent B cell clones, whereas in the two other siblings a monoclonal rearrangement pattern was found. All four patients showed clonal chromosome aberrations, which were different in each patient. In the two patients with biclonal Ig gene rearrangement patterns, two unrelated clones could also be demonstrated by the cytogenetic studies. These combined Ig gene and cytogenetic data indicate the presence of two different B-CLL in two of the four patients. Remarkably, the B-CLL cells of the two oldest patients expressed the CD8 antigen, which is rarely observed. Our finding of six different B-CLL in the four living siblings indicates that the members of this family are highly susceptible to the development of B-CLL. However we could not identify a common factor to explain this susceptibility further. In contrast to the literature, the occurrence of two B-CLL in one patient and the expression of CD8 were not associated with clinically aggressive disease in this family.

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Section: Southern Blot Analysis Of Ig Genesmentioning

confidence: 99%

Section: Southern Blot Analysis Of Ig Genesmentioning

confidence: 99%

Section: Southern Blot Analysis Of Ig Genesmentioning

confidence: 99%

Section: Southern Blot Analysis Of Ig Genesmentioning

confidence: 99%

Section: Southern Blot Analysis Of Ig Genesmentioning

confidence: 99%

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Four aged siblings with B cell chronic lymphocytic leukemia

F¹,

Dinkelaar

Hagemeijer

et al. 1997

Leukemia

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Detection of T-cell receptor delta gene rearrangement in T-cell malignancies by clonal specific polymerase chain reaction and its application to detect minimal residual disease

et al. 1996

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A clonal-specific polymerase chain reaction technique to detect T-cell receptor delta gene rearrangement in acute lymphoblastic leukaemia (ALL) and non-Hodgkin's lymphoma (NHL) was evaluated. It was applied to detect minimal residual disease. A sensitive and specific technique to detect minimal residual disease for T-cell malignancies was explored. Southern analysis and polymerase chain reaction (PCR) were used to detect the rearranged V-D-J segment of T-cell receptor delta (TCRG) gene from malignant cell specimens of patients with leukemia and lymphoma of T-cell lineage. The PCR product was sequenced and from the DNA sequences of the V-D-J region, a 3' anti-sense primer was designed and synthesized for clonal specific PCR (CS-PCR). T-cell receptor delta (TCR-6) gene rearrangement was studied in 40 cases of acute leukaemia and lymphoma of Tcell lineage at diagnosis. Using Southern analysis, the positive rates were 28 and 32% for the 18 T-lymphoma and 22 T-ALL, respectively. A one stage Polymerase Chain Reaction (PCR) technique was used to detect the rearrangement in Southern positive cases and the PCR positive rates were 80 and 86%, respectively. The PCR technique had a sensitivity of 0.1%. Serial follow-up marrow specimens were available from 4 T-ALL patients following chemotherapy for monitoring of minimal residual disease. Their PCR products were DNA sequenced. A 3' primer was designed for each case for a clonal specific (CS) PCR. The technique had a sensitivity of 0.003%. It was applied to detect minimal residual disease in serial follow-up marrow samples. The first patient had persistent negative CS-PCR results and enjoyed continuous remission for more than 3 years. The second patient with negative one stage PCR but positive CS-PCR results had eventual relapse of leukaemia. The other two patients never achieved a morphological remission. These preliminary results appeared to support the usefulness of these PCR techniques in detecting minimal residual disease and predicting relapses for ALL. However, further clinical correlation in larger populations of patients is necessary.

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