Quantification Of Leucocytes Research Articles

Quantification of leucocytes, T-lymphocytes and macrophages in autoptical endomyocardial tissue from 56 normal human hearts during the first year of life

This study evaluated the normal number of inflammatory cells in the heart in the first year of life using two methods to compare their ability to quantitate physiological myocardial infiltration. Eight endomyocardial samples from both ventricles were obtained at autopsy from 56 structurally normal hearts during the first year of life. In each sample the numbers of leucocytes, T-lymphocytes and macrophages were counted once in 20 randomly chosen high-power fields (400×) as well as in a 10mm2 area of randomly chosen myocardial tissue (100×) by two independent investigators. Compared to the literature a greater representative proportion of myocardial tissue was analyzed. The results of the enumeration in mm2 were converted into high-power-fields to compare both methods. The mean numbers and standard deviations for leucocytes, T-lymphocytes and macrophages were calculated. Both counting methods showed similar results with low inflammatory cell counts per single heart and staining. A greater understanding of the physiological myocardial infiltration by leucocytes, T-lymphocytes and macrophages is important for postmortem forensic cases, and for the interpretation of endomyocardial biopsies in infants.

New TiAg composite coating for bone prosthesis engineering shows promising microvascular compatibility in the murine dorsal skinfold chamber model.

The incorporation of antimicrobial substances like silver into implant surface coatings is one promising concept against primary infections of endoprosthesis, especially for immunocompromised patients as well as against reinfection after revision operations. However, besides good antimicrobial and mechanical properties it is equally important that the implant material does not disturb the local microvascular perfusion of muscle tissue to enable microbial host defense and tissue repair processes. In this study the biocompatibility of a newly developed TiAg-composite coating applied on conventional titanium via physical vapor deposition was analysed. To evaluate the local microvascular and inflammatory response of striated muscle tissue upon implantation of TiAg-coated plates the murine dorsal skinfold chamber model was used. We repetitively examined local capillary and venular perfusion, endothelial integrity as well as leucocyte activation by intravital fluorescence microscopy at 1 h, 24 h as well as 3 and 7 days after implantation. TiAg-implants were well tolerated by the vascular system as indicated by intact functional capillary density and endothelial integrity compared to pure titanium plates and controls without a metal implant. Furthermore, quantification of rolling and adherent leucocytes did not reveal signs of inflammation upon TiAg-implantation.

A comparative immunopathological study of injection site reactions in salmonids following intraperitoneal injection with oil-adjuvanted vaccines

An adequate understanding of the inflammatory mechanisms following intraperitoneally-injected oil-adjuvanted vaccines in fish is required for the improvement of the quality of vaccines. We report the findings of a study in which the importance of neutrophils, macrophages, lymphocytes and mast cells (MCs) in early inflammatory reactions of Atlantic salmon were examined in comparison with those of rainbow trout and grayling. The inflamed areas in the pylorus/pancreas area (injection site) were sequentially measured using computer-assisted microscopy, parallel with the quantification of leucocytes from 2 to 16 weeks post-vaccination. Antigen retention was also evaluated. Results show that inflammation in Atlantic salmon was slow to develop and was associated with a prolonged initial dominance of neutrophils. In contrast, the onset in rainbow trout was rapid and strong with early macrophage domination. The inflammatory and cellular response in grayling was initially in-between but progressed to be the strongest at 16 weeks. The results suggest that slow development of inflammation in Atlantic salmon and grayling leads to delayed sequestration of antigens resulting in chronically active inflammation that attempts to remove the antigens from the injection site. The strong onset in rainbow trout leads to rapid breakdown and removal of antigens resulting in early resolution.

Suspected SIDS and diagnosis of myocarditis

During the last years immunohistochemical techniques have improved the diagnosis of myocarditis using specific markers for quantification of leucocytes and characterization of T-lymphocytes together with the definition of MHC class II Antigens (HLA-DP, DQ, DR), known to be enhanced in cases of myocarditis. These techniques allow the diagnosis of myocarditis in an early stage of the inflammatory process in contrast to the conventional diagnostic with hematoxylin-eosin. 20 autopsy cases were examined in a retrospective study with conventional histological and immunohistological methods. After routine investigations including toxicological and histological examination of internal organs all cases were considered as cases of Sudden Infant Death Syndrome (SIDS). Increased number of LCA-positive leucocytes and CD-3-positive T-lymphocytes together with enhanced expression of inflammatory marker leads to the diagnosis of a lymphomonocytic myocarditis in three cases, in one case myocarditis could be diagnosed by conventional hematoxylin-eosin staining. In one of these four cases of myocarditis a positive immunohistochemical reaction was found using a new antibody against enteroviruses. Immunohistochemical techniques improve diagnosis of myocarditis in cases of suspected sudden infant death syndrome. Further studies using immunohistochemical inflammatory markers to control the incidence of acute myocarditis are necessary.

Immunohistochemical techniques improve the diagnosis of myocarditis in cases of suspected sudden infant death syndrome (SIDS)

Immunohistochemical techniques have improved the diagnosis of myocarditis. In a post mortem study, eight specimens in each case of the formalin-fixed and paraffin-embedded hearts of 20 suspected cases of sudden infant death syndrome (SIDS) were investigated with traditional hematoxylin–eosin staining and immunohistochemical methods. The hematoxylin–eosin stained specimens were examined for myocarditis according to the Dallas criteria; only in one case was a myocarditis diagnosed. The subsequent definition of the major histocompatibility complex class II antigens (HLA-DP,DQ,DR and HLA-DR), known to be enhanced in cases of myocarditis, the quantification of leucocytes with leucocyte common antigen (LCA) and characterization and quantification of T-lymphocytes using a specific marker (CD-3) allowed the definite diagnosis of myocarditis in three additional cases, six cases were found with moderate changes and ten cases without signs of inflammation.

Interstitial activated (IL-2R+) mononuclear cells and la antigens in experimental focal glomerulosclerosis

Tubulointerstitial localization of major histocompatibility complex (MHC) class II (Ia) antigens and immune-activated mononuclear cells (IL-2R+) and its suppression by prednisolone (PSL) were studied in a progressive model of focal glomerulosclerosis (FGS) induced by aminonucleoside (AN) and protamine sulfate (PS) given as 4 four day courses in rats. Tissues were prepared for the quantification of leucocytes on days 24, 52 and 80. The numbers of total leucocytes (CD45+), Ia+ cells, monocytes/macrophages, T lymphocytes (CD5+) and IL-2R+ cells (CD25+) were enumerated in immunoperoxidase sections using monoclonal antibodies: OX1, OX6, ED1, OX19 and ART18, respectively. The effects of PSL at a dose of 3 mg/kg body weight, which was begun from day 30, 12 days after the second series of injections, and continued for 50 days, were studied on day 80. There was no significant increase of glomerular T cells or IL-2R+ cells throughout the study. By contrast, each cell group in the interstitium increased in number throughout the disease evolution. Specifically IL-2R+ cells, which were rarely seen before AN + PS injection, were expressed by 5% of total leucocytes on day 80. Ia antigens also increased time-dependently and were found in proximal tubular cells on day 80. PSL treatment suppressed IL-2R and Ia expression in proportion to the decrease of total leucocytes and T cells. These results imply that delayed type hypersensitivity related to the appearance of immune activated T cells (IL-2R+) and MHC class II antigens may play a role in the progression of interstitial lesions and consequent renal dysfunction in a nonimmune-initiated FGS model.

Enzymatic quantification of polymorphonuclear leucocytes in normal and psoriatic skin following standardized injury.

Summary The intra-epidermal accumulation of polymorphonuclear leucocytes (PMN) following sellotape stripping of psoriatic non-lesional skin and normal skin of healthy individuals has been quantified with the elastase assay. Far more PMN accumulated in the skin in unstable psoriasis than in stable psoriasis. No significant difference was found, however, in PMN accumulation between the psoriatics as a whole and the controls. PMN accumulation was followed by the induction of an elastase inhibitor in psoriatic patients and controls. No significant differences could be found in inhibitory activity between controls and psoriatics. Our results suggest that the degree of PMN accumulation following surface trauma is essentially normal in stable chronic plaque psoriasis, but increases in unstable phases of the disease.