Adult Swiss (susceptible) and BALB/c (non-susceptible) mice were inoculated by the intravenous route with 1 × 106 yeast cells of Paracoccidioides brasiliensis, strain 18 Immunologic parameters, histopathology and features of the bronchoalveolar lavage (BAL) were evaluated at week 2, 4, 8 and 16 post-infection The pulmonary infection was progressive in Swiss mice and regressive in BALB/c mice The numbers of total cells, lymphocytes and polymorphonuclear neutrophils increased in BAL, as well as the percentages of giant cells, and CD4 and CD8 positive cells The ultrastructural study of BAL cells revealed a predominance of macrophages and a frequency of 132% of type II pneumocytes As the infection progressed, the number of fungal cells and spreading macrophages, as well as the stimulated release of H2O2 by macrophages, increased The animals exhibited an exacerbation of the humoral immune response and a depression of cellular immunity during the infection There was a good correlation between the intensity and the pattern of the pulmonary histopathology and the cellular findings in the BAL The present model reproduces some anatomoclinical patterns of the human disease and shows that BAL may be a useful tool in monitoring the pulmonary infection caused by P brasiliensis

Study of pulmonary experimental paracoccidioidomycosis by analysis of bronchoalveolar lavage cells: Resistant vs. susceptible mice

Objective To identify potential diagnostic markers of lupus nephritis (LN) based on bioinformatics and machine learning and to explore the significance of immune cell infiltration in this pathology. Methods Seven LN gene expression datasets were downloaded from the GEO database, and the larger sample size was used as the training group to obtain differential genes (DEGs) between LN and healthy controls, and to perform gene function, disease ontology (DO), and gene set enrichment analyses (GSEA). Two machine learning algorithms, least absolute shrinkage and selection operator (LASSO) and support vector machine-recursive feature elimination (SVM-RFE), were applied to identify candidate biomarkers. The diagnostic value of LN diagnostic gene biomarkers was further evaluated in the area under the ROC curve observed in the validation dataset. CIBERSORT was used to analyze 22 immune cell fractions from LN patients and to analyze their correlation with diagnostic markers. Results Thirty and twenty-one DEGs were screened in kidney tissue and peripheral blood, respectively. Both of which covered macrophages and interferons. The disease enrichment analysis of DEGs in kidney tissues showed that they were mainly involved in immune and renal diseases, and in peripheral blood it was mainly enriched in cardiovascular system, bone marrow, and oral cavity. The machine learning algorithm combined with external dataset validation revealed that C1QA(AUC = 0.741), C1QB(AUC = 0.758), MX1(AUC = 0.865), RORC(AUC = 0.911), CD177(AUC = 0.855), DEFA4(AUC= 0.843)and HERC5(AUC = 0.880) had high diagnostic value and could be used as diagnostic biomarkers of LN. Compared to controls, pathways such as cell adhesion molecule cam, and systemic lupus erythematosus were activated in kidney tissues; cell cycle, cytoplasmic DNA sensing pathways, NOD-like receptor signaling pathways, proteasome, and RIG-1-like receptors were activated in peripheral blood. Immune cell infiltration analysis showed that diagnostic markers in kidney tissue were associated with T cells CD8 and Dendritic cells resting, and in blood were associated with T cells CD4 memory resting, suggesting that CD4 T cells, CD8 T cells and dendritic cells are closely related to the development and progression of LN. Conclusion C1QA, C1QB, MX1, RORC, CD177, DEFA4 and HERC5 could be used as new candidate molecular markers for LN. It may provide new insights into the diagnosis and molecular treatment of LN in the future.

/pdf/predicting-diagnostic-gene-expression-profiles-associated-2svjtzdn.pdf

Predicting diagnostic gene expression profiles associated with immune infiltration in patients with lupus nephritis

We investigated the reaction of the cellular immune system of liver and blood in the C57BL/6 mouse to a metastasizing Lewis lung carcinoma. The cellular immune system of the liver consists of mature and immature macrophages, B-cells, T-cells including their subpopulations, and natural killer cells, and their percentage frequencies differ significantly from those in the corresponding mononuclear blood cell (MBC) compartment. This suggests that the hepatic immune cells represent a system with autonomous function showing a typical homing of its members. Imminent metastasis to the liver is signalled by impressive alterations in the percentage frequencies of nonparenchymal liver cells (NPLC). There are a dramatic loss of mature macrophages, an increase in immature macrophages, a reduction of T-helper cells leading to a low CD4/CD8 ratio, and an increase in natural killer cells. In the blood, the corresponding precursor cells show comparable changes with a delay of at least 2 days. Early metastasis is accompanied by a significant increase in mononuclear NPLC producing tumour necrosis factor alpha. The alterations in percentage frequencies of the NPLC during tumour metastasis differ markedly from the changes in these cells in the liver during endotoxinaemia.

Early detection of metastasis by alterations in the cellular immune system in the murine liver and blood.

The object of the present investigation was to extend fundamental understanding of the composition, function and origin of bronchoalveolar lavage (BAL) cells, and to reduce the considerable gaps in our existing knowledge. BAL cells from the B10.D2 mouse were identified by morphological, enzyme cytochemical and immunocytochemical methods and a further functional characterisation of macrophages was undertaken in vitro using latex phagocytosis. The possible bone marrow origin of BAL cells was investigated with a radiation chimera. The bone marrow from H2k-positive F1 mice (B10.BR×B10.D2) was transplanted into the irradiated H2k-negative parent animals. At various time intervals after transplantation, BAL cells were tested immunocytochemically for the presence of H2k-positive cells. The principal finding was that BAL cells are functionally heterogeneous. It was possible by morphological, enzyme cytochemical and immunocytochemical techniques to subdivide the total BAL cell count into 89% macrophages, 8.5% lymphocytes and 2.5% granulocytes. Only 14.3% of the macrophages expressed the corresponding epitope, and this small population could be further subdivided immunocytochemically into mature and immature (7∶1) macrophages. The ability to phagocytose, which is typical of active macrophages, could be induced in vitro in only one-third of all BAL macrophages. Three-quarters of the macrophages carried the H2d antigen and 9.5% of the BAL cells were natural killer cells showing a macrophage phenotype. Subdivision of the lymphocytes showed a significant majority of T-cells (75%) to B-cells (20%), and 5% Asialo GM1-positive cells. The CD4/CD8 ratio amounted to 1.3∶1 and 38% of the lymphocytes expressed on the surface the H2d antigen. Throughout the experiment with the chimeras (6 to 52 weeks), H2k-positive BAL cells could be detected. These observations verify the participation of the bone marrow in the replacement of available intraalveolar cells. These findings show that the lung, as indicated by BAL cells, has at its disposal a highly developed mobile cellular defence mechanism, of which the numerically dominant component is the macrophage population. These cells should be regarded as possessing various as yet largely unknown functions.

Function and "homing" of the lung macrophages - I. Evidence of functional heterogeneity of mobile cells of the murine lung parenchyma in the bronchoalveolar lavage (BAL)

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Function and "homing" of the lung macrophages - I. Evidence of functional heterogeneity of mobile cells of the murine lung parenchyma in the bronchoalveolar lavage (BAL)

Early detection of metastasis by alterations in the cellular immune system in the murine liver and blood.