Establishment and characterization of hybrid rat mast cells

Zheng, Yunli; Chan, Bosco M.C.; Rector, Edward S.; Berczi, István; Froese, Arnold

doi:10.1016/0014-4827(91)90369-6

Cited by 11 publications

(20 citation statements)

References 26 publications

(1 reference statement)

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“…The RBL-hEIa-2B12 cell line, which has a mucosal mast cell phenotype [16], was transfected with the α-chain of human Fcε receptor type 1 (huFcεR1) complex (a kind gift from Dr. Teshima, Japan) [17]. Degranulation of RBL-hEIa-2B12 cells with pooled serum from CMA patients was performed as described previously [18].…”

Section: Methodsmentioning

confidence: 99%

Enzymatic Treatment of Whey Proteins in Cow’s Milk Results in Differential Inhibition of IgE-Mediated Mast Cell Activation Compared to T-Cell Activation

Knipping

Esch

Dijk

et al. 2012

Int Arch Allergy Immunol

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Background: Cow’s milk (CM) hydrolysates are frequently used as milk substitutes for children with CM allergy. In hydrolysates, allergenic epitopes within CM proteins are diminished by enzymatic treatment. The aim of this study was to examine the allergenic and immunogenic properties of whey proteins during hydrolysis. Methods: During hydrolysis, samples were obtained at 0, 10, 15, 30, 45, 60, 75 and 90 min. Degradation was checked by HPLC and SDS-PAGE. Allergenic potential was analyzed by IgE crosslinking capacity of human Fcε receptor type 1-transduced rat basophilic leukemia cells sensitized with serum of CM-allergic patients. Whey-sensitized C3H/HeOuJ mice were ear challenged intracutaneously with the hydrolysates. Immunogenicity was tested using whey-specific human T-cell clones and T-cell lines at the level of proliferation and release of IL-4, IL-10, IL-13 and IFN-γ. Results: After 15 min of hydrolysis, the majority of the proteins were degraded. Hydrolysis for 15 min resulted in 92% inhibition of mast cell degranulation and in 82% reduction of ear swelling in the mouse model. In contrast, T-cell-stimulatory capacity was less affected by hydrolysis: reduction of human T-cell proliferation was only 9%. This was further reduced to 57 and 74% after 30 and 45 min of hydrolysis, respectively. Cytokine production followed the pattern of T-cell proliferation. Conclusion: Via differential analysis of allergenic versus immunogenic properties of the time kinetics of hydrolysis of whey proteins, we have demonstrated specific hydrolysis conditions with reduced IgE-crosslinking responses but retained T-cell activating properties. This approach might be useful in better defining CM hydrolysates.

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Section: Methodsmentioning

confidence: 99%

Enzymatic Treatment of Whey Proteins in Cow’s Milk Results in Differential Inhibition of IgE-Mediated Mast Cell Activation Compared to T-Cell Activation

Knipping

Esch

Dijk

et al. 2012

Int Arch Allergy Immunol

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“…The present study was undertaken in order to confirm that the presence of hybrid mast cells with the CTMC phe notype was not an isolated event, since in the previous study [8] only few such cells were found in one of the primary cultures. Moreover, it was considered important to confirm the presence of CTMC-like hybrid cells by monitoring more than one phenotypic parameter and to further explore the possibility that the disappearance o f the CTMC pheno type was a consequence of chromosome loss.…”

Section: Introductionmentioning

confidence: 99%

“…It was established and maintained as described previously [8]. Hybrid rat mast cells (HRMC) were maintained in RPMI 1640 medium (Gibco, Grand Island, N.Y., USA) supplemented with 10% fetal calf serum and other nutrients as suggested by the manufacturer.…”

Section: Cells and Cell Culturementioning

confidence: 99%

“…Fusion of 6-thioguanine-resistant RBL-CK2-TG cells and RPMC was essentially carried out as described previously [8], except that polyethylene glycol of molecular weight 1,500 (Merck, Darmstadt, Germany) was used and the fusion was carried out in scrum-free RPMI medium. Once cell growth was established, the primary cultures were immediately expanded and aliquots were frozen for storage in liquid nitrogen.…”

Section: Hybridization O F Rpmc and Rbl Cellsmentioning

confidence: 99%

“…Rast mast cells are represented by two phenotypically distinct cell types: the connective tissue type (CTMC) and the mucosal type (MMC) [1,2], Rat peritoneal mast cells (RPMC) are among the most common members o f the CTMC family [3] while rat basophilic leukemia (RBL) cells are frequently used as representatives of the MMC pheno type [4], They originate in the bone marrow [2,5] and their growth and differentiation is regulated by microenviron mental factors, including several interleukins and stem cell factor [5][6][7] We have recently reported the establishment of hybrid rat mast cells (HRMC) [8], They were generated by the fusion of RPMC and HAT (hypoxanthine, aminopterin, thymidine)-sensitive RBL cells in the presence o f polyethylene glycol [8], Only relatively few cells o f the CTMC pheno type could be detected among the hybrid cells by differen tial staining using alcian blue and safranin O. Attempts to confirm the presence o f cells of this phenotype by other methods failed.…”

Section: Introductionmentioning

confidence: 99%

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Phenotypic Changes among Hybrid Rat Mast Cells

Zheng

McNeill²,

Rector³

et al. 1995

Int Arch Allergy Immunol

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Rat peritoneal mast cells and 6-thioguanine-resistant rat basophilic leukemia cells, representative of connective tissue-type (CTMC) and mucosal (MMC) mast cells, respectively, were fused using polyethylene glycol. Four out of 14 primary hybrid mast cell lines contained more than 50% of CTMC as demonstrated by histochemical staining. Two cell lines, one predominantly of the CTMC and the other of the MMC phenotype, were selected for further study. Among these, the phenotype was also confirmed by analysis for rat mast cell protease I and by mediator release triggered by compound 48/80 and ionophore A23187. The CTMC phenotype disappeared after culturing cells for 2 weeks. The change in phenotype did not significantly alter the mediator release due to calcium ionophore A23187. Repeated cloning of cells bearing the CTMC phenotype did not yield a cloned line of cells expressing the CTMC phenotype only, although it prolonged the persistence of this phenotype. During the period of CTMC phenotype loss, a drop in cellular DNA content occurred, suggesting that chromosome instability may, at least partially, have been responsible for the phenotypic changes.

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