Mouse × human heterohybridomas as fusion partners with human B cell tumors

Variations in “rescuability” of immunoglobulin molecules from different forms of human lymphoma: implications for anti-idiotype vaccine development

“…Loss of Id secretion by growing hybridomas accounted for only 3/18 cases of failure to produce Id vaccine. Interestingly, the Id vaccine production success rate was substantially lower in cases of MCL, in contrast to what was initially described with the very same method [16] in newly diagnosed MCL patients. Indeed, researchers at the NCI have recently reported that the use a rescue fusion-related Id vaccine for MCL patients is feasible in 96% of cases [20].…”

“…As the same fusion partner (K6H6/B5) was used with all the different malignant B-cell tumor cells [16], these results seem to imply that B-cells other than those constituting FL clonal populations are less prone to originate suitable hybridomas with the same fusion method utilized for FL Id vaccine production. This conclusion is further supported by the fact that only one fusion attempt was required in most FL cases, as opposed to the other NHL subtypes where the pre-established, arbitrary, five-attempt cut-off, was reached in most instances, irrespective of the ultimate Id vaccine production outcome.…”

“…A large number of tumor cells are required for the fusion with a purposely-selected cell line (K6H6/B5, i.e. ATCC CRL 1823) according to the published methods [16]. Among the several hybridomas grown (Fig.…”

“…Inclusion was based on an attempt to produce an Id vaccine only at the time of first relapse and on the previous, formal demonstration of the presence of a complete, clonal and tumor-specific immunoglobulin on the tumor cell surface. The diagnosis at relapse was always confirmed pathologically and Id rescue was always carried out according to the traditional hybridoma technique laid out above [16].…”

Generation and Characterization of two Monoclonal Self‐Associating IgG Rheumatoid Factors from a Rheumatoid Synovium

“…Mononuclear cells (SFMC) were isolated from these samples and were cultured in growth medium (GM) (RPMI 1640 supplemented with 10% fetal bovine serum, 1% nonessential amino acids, 1 mM sodium pyruvate, 10 mM HEPES, 100 units/ml penicillin, 100 pg/ml streptomycin, and 2 mM L-glutamine) at 37°C in a humidified 5% CO, incubator for 3 days. Approximately 4 x 10" cultured SFMC were mixed with 2 x 10" K6H6/BS cells (7,9), and the suspension was centrifuged at 250g for 5 minutes at room temperature (RT). After removing the supernatant, the cell pellet was gently resuspended in 1 ml of RPMI 1640 with 35% (weightholume) polyethylene glycol.…”