The coagulum consists of the whole Casein fraction (., the solid fraction of proteins obtained after coagulation of Milk). It is subdivided into a number of families, of which the most important are a S1 -, a S2 -, b -, K-, Y-caseins (alphaS1 -, alphaS2 -, beta-, kappa-, gamma-caseins) (4). Each individual Casein represents a well-defined chemical compound, but they cross-link to form ordered aggregates (nanoclusters) (., micelles) that assemble into larger structures, forming Casein micelles characterised by a central hydrophobic part and a peripheral hydrophilic layer in suspension in Lactoserum (Whey) (13-14). Their proportion in the micelles is relatively constant at approximately 37%, 13%, 37%, and 13%, respectively.
The main characteristics that should be emphasised are the multiplicity and diversity of proteins that are involved in Cow’s milk allergy (CMA). Polysensitisation to several proteins occurs most often, and all Milk proteins appear to be potential allergens (1). A great variability is observed in the affinity, specificity and magnitude of IgE responses in patients’ sera (15). Most Milk-allergic patients are sensitised to several proteins, including BLG (Bos d 5), Casein (Bos d 8), ALA (Bos d 4), BSA (Bos d 6), Lactoferrin, and Immunoglobulins (Bos d 7) (10,14,16-22). A great variability is observed in IgE response (1).
Casein and BLG, as well as ALA, are major allergens. However, proteins present in very low quantities, such as BSA, immunoglobulins, and especially lactoferrin, also appear to be important since 35% to 50% of patients are sensitized to those proteins and sometimes to those proteins only (19). In the last few years, sensitivity to Casein seems to have increased in terms of both frequency and intensity of IgE response (1). Sensitizations to Casein, BLG, and ALA are closely linked. In contrast, sensitivity to BSA appears to be completely independent, with 50% of the patients being sensitized to BSA regardless of their sensitivity to other Milk allergens (1).
The role of various Cow’s milk proteins (CMPs) in the pathogenesis of CMA is still controversial. Sera from 20 Milk-allergic subjects have been used for Cow’s milk major allergen identification. The prevalence of CMP allergens has been measured as the following: 55% Alpha(s1)-casein, 90% Alpha(s2)-casein, 15% Beta-casein, 50% Kappa-casein, 45% Beta-lactoglobulin, 45% BSA, 95% IgG-heavy chain, 50% Lactoferrin, and 0% Alpha-lactalbumin (23).
Allergens from Bos domesticus listed by IUIS*
LLC-PK1 and L-MDR1 cells are seeded in 96-well culture plates with microporous polycarbonate inserts and grown for 4 days in the same medium as used for cell cultures but without vincristine. The medium is replaced every 2 days. Before running the assay, the culture medium is replaced by HBSS buffer supplemented with 10 mM HEPES. Rivaroxaban are dissolved in DMSO and diluted with transport buffer to the respective final test concentrations (final DMSO concentration is always 1%). For inhibitor studies, the inhibitor is added at the appropriate concentration. counted. After 2 hour incubation at 37 °C, samples are taken from both compartments and, after the addition of ammonium acetate buffer and acetonitrile, are analyzed by LC-MS/M