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The central 95 percentile estimates of the normal white cell types (as determined by a standard differential count) were calculated from 777 normal individuals. The results were divided into groups by age and sex and expressed both as percentages and as absolute numbers of cells.A similar survey was made using the prototype of a new automated differential counting method (Hemalog-D(TM)). The two sets of laboratory values showed no statistically significant differences regarding age or sex and were strikingly similar in most cell types with the exception of band and juvenile forms of granulocytes. As expected, the normal range using the manual technique was somewhat broader than that found with the automated method. In addition to being the first comparison of machine counts with visual differentials this study provides additional insight into the age distributions of normal cells.

Guinea pig eosinophil granules are characterized by the presence of a basic protein of low molecular weight which accounts for greater than 50% of granule protein. This protein, termed the major basic protein (MBP), readily aggregates and becomes insoluble, and the formation of aggregates is dependent on the establishment of disulfide bonds. Analysis of concentrated preparations of MBP often revealed a series of bands which were multiples of a monomeric unit with a mol wt of approximately 11,000. Analysis of reduced and alkylated MBP on a 10% agarose column equilibrated with 6 M guanidinium chloride revealed a single polypeptide chain with a mol wt of 10,800. Amino acid analysis of the protein revealed the presence of 13% arginine, consistent with the basic character of the molecule. Four residues of tryptophan, were present, indicating that MBP is not a histone. The MBP did not increase vascular permeability when injected into the skin of guinea pigs, nor did it antagonize the effect of histamine and bradykinin in the skin. MBP also did not contract the isolated guinea pig ileum and when mixed with histamine or bradykinin did not inhibit their activity on the gut. MBP had only weak, if any, antihistaminic activity. MBP possessed weak bactericidal activity when compared to histone and then only with one strain of E. coli. MBP precipitated DNA, neutralized heparin, and activated papain. On a molar basis MBP was more active than cysteine in activating papain. These results do not point to any unique biological activity associated with MBP other than those expected of a protein as basic as it is and one which possesses reactive sulfhydryl groups. Possible functions of eosinophils based on the properties of the MBP are discussed.

Charcot-Leyden crystals develop spontaneously in certain diseases of man and can be formed within minutes from eosinophils lysed with a surface-active agent (Aerosol-OT). Treated human eosinophils examined by electron microscopy showed the general features of cell lysis. After disruption of the eosinophilic granule of man, there remained an insoluble crystalline core, which was now more dispersed, a few tubular structures resembling microtubules and a fine granular accumulation around the periphery of the granule. It is suggested that such dispersion represents rearrangement of the crystalline structure allowing the material of the core to be incorporated into a Charcot-Leyden crystal. In guinea pig eosinophils, Charcot-Leyden crystals were not found even after prolonged lysis. By electron microscopy, the cellular changes in guinea pig eosinophils were generally similar to, but developed more slowly than, those in human eosinophils. Tubular structures detected in the cortical region of the granule were more numerous than thosenoted in the disrupted eosinophil granule of man. The different reaction to injury of the eosinophils of man and those of the guinea pig appears to be characteristic for each of the two species.