Comparison of methods of counting skin window coverslips A statistical analysis

Garrie, Stuart A.; Wolf-Jürgensen, P

doi:10.1016/0091-6749(72)90086-3

Cited by 3 publications

(3 citation statements)

References 9 publications

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“…experiments, cover slips were removed from Petri dishes or Leighton tubes, dipped twice in physiologic saline, fixed in Helly's solution (Zenker-formol), stained with May-Grtinwald-Giemsa by the method of Jacobson and Webb (18), and mounted for microscopic examination. The coverslips were coded and counted by one investigator with a fixed method of scanning (19). Generation times in fibroblasts and peritoneal cells were calculated from cover slip counts allowing for error due to multiple infection (20).…”

Section: Measurement and Analysis Of In Vitro Assays--at Each Intervmentioning

confidence: 99%

Cell-Mediated Immunity Against Besnoitia and Toxoplasma in Specifically and Cross-Immunized Hamsters and in Cultures

Hoff

Frenkel

1974

The Journal of Experimental Medicine

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The capacity of hamster peritoneal cell populations to control viability and growth of Besnoitia and Toxoplasma organisms was assessed in vivo and in vitro. Immunized hamsters reduced the homologous organisms 100- to 10,000-fold over a 5-day period, but the heterologous infection increased 100- to 1,000-fold in numbers, similar as in the nonimmune controls. Passively administered antibody was ineffective although lytic cofactors were supplied by hamsters. In cultures, peritoneal cells from Besnoitia-immune hamsters delayed the growth of homologous parasites to an average of 38.5 h per division; however, in Toxoplasma-immune and nonimmune cells, Besnoitia divided every 12.8 h. Specificity of immunity was pronounced against both infections. With cross-infections, Toxoplasma-immune cultures did not effectively delay Besnoitia growth; however, Besnoitia-immune cultures reduced Toxoplasma growth by one-half. Co-cultivation experiments demonstrated that specifically committed lymphocytes could instruct macrophages to reduce the homologous organism 10-fold, whereas heterologous organisms were reduced only 2-fold. Lymphocyte supernatants initiated hypersensitivity as indicated by macrophage activation and giant cell formation in culture. However, these supernatants did not transfer infection immunity. Lymphokines could account for the hypersensitivity phenomena, but cell-mediated infection immunity in this model required close lymphocyte-macrophage proximity. These studies indicate that a number of distinct processes including delayed hypersensitivity, macrophage activation, and specific cellular immunity are acting simultaneously during latent Besnoitia infection of hamsters. All three processes are mediated by lymphoid cells and appear to be specifically induced. Although activated macrophages develop some heightened nonspecific capabilities, these were several orders of magnitude below the specific effects.

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Section: Measurement and Analysis Of In Vitro Assays--at Each Intervmentioning

confidence: 99%

Cell-Mediated Immunity Against Besnoitia and Toxoplasma in Specifically and Cross-Immunized Hamsters and in Cultures

Hoff

Frenkel

1974

The Journal of Experimental Medicine

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“…Slides were viewed under oil power objective (1,OOOX) and cells were counted by a "fixed method" (11) in which a predetermined and fixed coordinate system was used to determine whether a cell was included in the differential. Usually 200-400 cells were counted per cover slip, but when only a few hundred cells were present, all cells on the entire coverslip were counted.…”

Section: Introductionmentioning

confidence: 99%

Basophils in tuberculin and "Jones-Mote" delayed reactions of humans.

Askenase¹,

Atwood²

1976

J. Clin. Invest.

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A B S T R A C T Jones-Mote reactions are delayed, erythematous, and mildly indurated cutaneous reactions originally described in humans sensitized by skin injection of heterologous proteins. Similar reactions in guinea pigs contain many basophils and are called cutaneous basophil hypersensitivity. In contrast, guinea pigs immunized with mycobacterial adjuvants have classical tuberculin-type delayed hypersensitivity reactions, which contain few basophils. This has led to a new classification of delayed responses, based largely on the presence or absence of basophils. We induced sensitization for JonesMote reactions in 20 normal humans by intradermal injections of keyhole limpet hemocyanin. Skin tests with KLH 1 wk later showed erythematous and indurated delayed reactions in all subjects. Rebuck skin windows showed specific accumulations of basophils with a delayed time-course in 18 of 20 subjects. In 12 normals sensitized with oxazolone-keyhole limpet hemocyanin conjugates, skin reactions and in vitro lymphocyte stimulation showed carrier and not hapten specificity, suggesting that cutaneous responses were probably mediated by T cells.A comparative stuidy of strongly positive PPD skin tests in patients with tuiberctulosis showed significant basophil accuimulations in five of nine subjects. Thus, basophils occuirred in human tuibercullin and Jones-Mote reactions and were not a distinguishing feature of Jones-Mote reactions. We suggest that the occurrence of basophils at delayed reactions is under complex regulation and that basophil accumulations are an aspect of delayed hypersensitivity, rather than an indication of a distinctive and separate response. Dr. Askenase is the recipient of Allergic DiseasesAcademic Award AI-70829, and Dr. Atwood was the recipient of a student fellowship from the Syntex Corporation.

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“…The coverslips were changed at 3, 6,9,12,15,21,24,27,30,33,36,39,45 and 48 h, air dried, and Wright stained. The cells were counted in a fixed manner (Garrie & Wolf-Jurgensen, 1972) and the percentage of each cell type calculated.…”

Section: Methodsmentioning

confidence: 99%

Asymmetry of cellular response as revealed by the skin window technique

Garrie

Crump

1973

Br J Dermatol

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summary Utilizing the skin window technique, symmetry of cellular response was examined by comparing cellular exudates from symmetrical areas of opposite forearms in eleven subjects. Testing the hypothesis that there would be no significant difference between symmetrical sites subjected to the same stimulus at the same time, the percentages of cell types in the cellular exudate were calculated for 126 paired comparisons. A significant difference in the cell types arriving at the skin window sites occurred in seventy‐six out of the 126 paired comparisons (Chi square analysis, P<0.01). This lack of a symmetrical response could be due to an inherent variability of the cellular response or to an inability to manipulate the two sites in exactly the same manner. Although significant differences were found between the above individual matched pairs (Chi square analysis), the ‘t’ test analysis of the pairs grouped by hour revealed no significant difference in mean percentages of cell types between left and right arms at fourteen time intervals. Thus, previous skin window investigations comparing mean percentages of cell types were found to be valid, even though symmetry of cellular response in individual subjects is much less than expected.

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Comparison of methods of counting skin window coverslips A statistical analysis

Cited by 3 publications

References 9 publications

Cell-Mediated Immunity Against Besnoitia and Toxoplasma in Specifically and Cross-Immunized Hamsters and in Cultures

Cell-Mediated Immunity Against Besnoitia and Toxoplasma in Specifically and Cross-Immunized Hamsters and in Cultures

Basophils in tuberculin and "Jones-Mote" delayed reactions of humans.

Asymmetry of cellular response as revealed by the skin window technique

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