Sistema monocítico-macrofágico y mycobacterium leprae: su importancia en la interpretación patogénica de las distintas formas clínico-patológicas de la Lepra y de las reacciones leprosas / Monocytic-macrophagic system and mycobacterium leprae

Se realiza una revisión de la patogenia de la Lepra (Hanseniasis), en sus vinculaciones con el "sistema monocítico-macrofágico y se efectúa el estudio histopatológico, histoquimico (lipídico) y con microscopía electrónica de cuatro enfermos de Lepra, dos lepromatosos ( sin y con eritema nudoso) y otros dos dimorfos ("Borderline"). Se concluye que: 1) La Lepra presenta sus manifestaciones clínico-patológicas más ostensibles, en vinculación con lesiones del "sistema monocítico-macrofágico". 2)Existen distintos comportamientos de los macrófagos frente al mycobacterium leprae" a)en personas Mitsuda positivos se puede demostrar el predominio de "macrófagos lisadores del M.leprae" con muerte y desaparición de los bacilos; b)en individuos Mitsuda-negativos predominan los "magrófagos no lisadores del M.leprae",que también producen la muerte de los bacilos, pero con persistencia de la "envoltura lipídica bacilar", que se deposita en las vacuolas de los virchowcitos; y c)en enfermos Mitsuda-negativos, luego de formado el "Granuloma virchowiano", se desarrollan los "macrófagos lisadores de las células de Virchow" que permitirían el recambio celular de los virchowitos. 3)Los "macrófagos lisadores del M.leprae" obtienen información antigénica del "bacilo aislado" que pueden transmitir al sistema de inmunidad celular, cuyos linfocitos T activados generan los "granulomas epitelioides con células de Langhans" (granulomas tuberculoides), de la Lepra tuberculoide. 4) Los "macrófagos no lisadores del M.leprae" no obtendrían información antigénica adecuada y no tendrían capacidad para estimular ningún sistema inmunológico (celular, o humoral), originando los "granulomas virchowianos" de la Lepra lepromatosa 5)Luego del envejecimiento y muerte de los vinchowcitos, los "macrófagos lisadores de las celulas de Virchow" obtendrían información antigénica de los "bacilos degenerados asociados a lípidos y glucoproteinas citoplasmáticas", con capacidad de estimulación del sistema inmunológico humoral cuyos linfocitos B activados generarían anticuerpos complejos: a)contra bacilos ubicados en los virchowcitos en la Reacción leprosa tipo 2 (eritema nudoso leprótico); b) contra la pared de vasos en la reacción leprosa tipo 3(fenómeno de Lucio); y c)contra tejidos del huésped (enfermedad autoagresiva hanseniana-Azulay). 6)En la lepra no existiría "polaridad inmunológica" en los períodos iniciales de la instalación de la enfermedad, pero parecería secundariamente a la formación del "granulo

Sistema monocítico-macrofágico y mycobacterium leprae: su importancia en la interpretación patogénica de las distintas formas clínico-patológicas de la Lepra y de las reacciones leprosas / Monocytic-macrophagic system and mycobacterium leprae

Se realiza una revisión de la patogenia de la Lepra (Hanseniasis), en sus vinculaciones con el "sistema monocítico-macrofágico y se efectúa el estudio histopatológico, histoquimico (lipídico) y con microscopía electrónica de cuatro enfermos de Lepra, dos lepromatosos ( sin y con eritema nudoso) y otros dos dimorfos ("Borderline"). Se concluye que: 1) La Lepra presenta sus manifestaciones clínico-patológicas más ostensibles, en vinculación con lesiones del "sistema monocítico-macrofágico". 2)Existen distintos comportamientos de los macrófagos frente al mycobacterium leprae" a)en personas Mitsuda positivos se puede demostrar el predominio de "macrófagos lisadores del M.leprae" con muerte y desaparición de los bacilos; b)en individuos Mitsuda-negativos predominan los "magrófagos no lisadores del M.leprae",que también producen la muerte de los bacilos, pero con persistencia de la "envoltura lipídica bacilar", que se deposita en las vacuolas de los virchowcitos; y c)en enfermos Mitsuda-negativos, luego de formado el "Granuloma virchowiano", se desarrollan los "macrófagos lisadores de las células de Virchow" que permitirían el recambio celular de los virchowitos. 3)Los "macrófagos lisadores del M.leprae" obtienen información antigénica del "bacilo aislado" que pueden transmitir al sistema de inmunidad celular, cuyos linfocitos T activados generan los "granulomas epitelioides con células de Langhans" (granulomas tuberculoides), de la Lepra tuberculoide. 4) Los "macrófagos no lisadores del M.leprae" no obtendrían información antigénica adecuada y no tendrían capacidad para estimular ningún sistema inmunológico (celular, o humoral), originando los "granulomas virchowianos" de la Lepra lepromatosa 5)Luego del envejecimiento y muerte de los vinchowcitos, los "macrófagos lisadores de las celulas de Virchow" obtendrían información antigénica de los "bacilos degenerados asociados a lípidos y glucoproteinas citoplasmáticas", con capacidad de estimulación del sistema inmunológico humoral cuyos linfocitos B activados generarían anticuerpos complejos: a)contra bacilos ubicados en los virchowcitos en la Reacción leprosa tipo 2 (eritema nudoso leprótico); b) contra la pared de vasos en la reacción leprosa tipo 3(fenómeno de Lucio); y c)contra tejidos del huésped (enfermedad autoagresiva hanseniana-Azulay). 6)En la lepra no existiría "polaridad inmunológica" en los períodos iniciales de la instalación de la enfermedad, pero parecería secundariamente a la formación del "granulo

Biphasic regulation of macrophage attachment by activators of cyclic adenosine monophosphate-dependent kinase and protein kinase C.

A method is described that enabled us to study the adhesiveness of J-774 murine macrophages. Cell attachment was stimulated by activators of kinase C (i.e., phorbol esters) as well as kinase A (cyclic adenosine monophosphate; cAMP). This novel effect of cAMP was observed when its levels were increased via receptor triggering (prostaglandin E1, beta-adrenergic agonists), activation of Ns (cholera toxin), or inhibition of phosphodiesterase (Ro 20-1724) or when the kinase was directly activated by Br8-cAMP. The simultaneous treatment with kinase A and kinase C activators at the time of attachment resulted in a partially additive response. On the other hand, preincubation of the cells in suspension with one of the activators rendered them refractory to subsequent stimulation at the onset of the adhesion assay, whatever agent was used. Such a refractoriness was also observed in cells preincubated with oleoyl-acetyl-glycerol (OAG). On the other hand, when added at the time of attachment, this near-physiological activator of kinase C evoked a biphasic response: the early stimulation of cell attachment was followed by an accelerated rate of "detachment." In conclusion, kinase C and kinase A play a role in the sequence of events leading to cell adhesion. The cross desensitization observed is distal and takes place at or beyond the kinase step.

Delayed maturation of skin window macrophages in myelodysplastic syndromes.

Blood monocyte differentiation to macrophages was examined in nine patients with primary myelodysplastic syndromes using the skin window technique. Emigrated cells were stained cytochemically for acid phosphatase reaction after 1, 2, 4, 7, 9, 12 and 23 h. Compared to age-matched controls, seven patients showed a significant delay in lysosomal enzyme acquisition, which is associated with macrophage differentiation. Our results with this in-vivo assay demonstrate an involvement of the monocyte/macrophage system in primary myelodysplastic syndromes and show that patients often have a disturbance in macrophage differentiation.

Inflammation of rat molar pulp and periodontium causes increased calcitonin gene-related peptide and axonal sprouting.

We have studied the response of nerve fibers containing calcitonin gene-related peptide immunoreactivity (CGRP-IR) to inflammation using a rate dental experimental system. Inflammation was induced by drilling tooth cusps to create pulpal exposures; the induced pulpitis and subsequent periapical lesions were studied 1-35 days later using standard CGRP immunohistochemistry and the avidin-biotin peroxidase method. The injury and resulting inflammation caused a disruption of CGRP-IR nerve fiber location and arborization that varied depending on whether the initial injury was limited to the pulp tip or extended throughout the pulp horn. At shorter survival periods (24 hr, 3 days) nerve fibers were either decreased or bundled into the center of the pulp with sprouting along the wound border. At 6 days necrosis and acute inflammation had advanced to varying degrees, and CGRP-IR fibers were extensively sprouted in the surviving pulp; the pulp also stained specifically for CGRP within 1-2 mm of the inflamed tissue at 6 days. At 35 days, we found total pulp necrosis in most teeth and the development of periapical bone loss, granulomatous tissue, and periapical abscesses. There was also an extensive increase in CGRP-IR nerve fibers in the tissues surrounding sites of severe periodontal inflammation and necrosis. In some cases, macrophage-like cells staining specifically for CGRP were near the abscesses. The results show important interactions between peptidergic nerve fibers and inflammatory cells, and are discussed in terms of the role of nerve fibers containing CGRP in neurogenic inflammation, mechanisms for intensification of CGRP immunoreactivity in affected fibers or neighboring cells, and implications for chronic inflammatory conditions, dental pain, and anesthesia.

Immortalization of cloned mouse splenic macrophages with a retrovirus containing the v-raf/mil and v-myc oncogenes.

The recombinant retrovirus J2, which contains the v-raf/mil and v-myc oncogenes, was used to immortalize mouse splenic macrophages that had been cloned in soft agar. When added to freshly harvested colonies, J2 failed to yield cell lines but it immortalized up to 30% of the clones if they had been maintained for at least 4 months in medium containing colony-stimulating factor 1 (CSF-1). All of the cell lines grew in agar in a CSF-1-independent manner, and they produced tumors in nude and syngeneic mice. The cell lines were judged to be macrophage based on morphological criteria and because they secreted lysozyme, were phagocytic for antibody-coated particles, and expressed both the Mac-1 antigen and the CSF-1 receptor. The cell lines could be divided into three groups based on their expression of Ia and their ability to present an antigen to a T-cell hybridoma. The majority of the lines did not constitutively express Ia or present antigen, but a lymphokine did induce Ia in all of the lines, with most of them also acquiring antigen-presenting activity. However, a small proportion of lymphokine-treated lines continued to lack antigen-presenting activity despite their ability to express Ia. The third and smallest group of cell lines constitutively expressed both Ia and antigen-presenting activity. These results show that the J2 recombinant retrovirus is a useful means of immortalizing functionally distinct populations of cloned splenic macrophages.

Isolation and characterization of glomerular macrophages in experimental glomerulonephritis.

Adherent cells emigrating from glomeruli of rabbits developing anti-glomerular basement membrane antibody induced glomerulonephritis were isolated and characterized as macrophages. Glomeruli were isolated using a sterile graded sieving technique and cultured in plastic tissue culture flasks. After varying culture times, emigrating adherent cells were harvested by 'cold shock' or trypsin-versene. These cells had the morphological and functional characteristics of macrophages. They were largely mononuclear, esterase-positive, phagocytic cells, which exhibited surface Fc receptors. A mean of 4.8 +/- 2.1 X 10(4) macrophages could be isolated from 2 X 10(4) glomeruli after 1 h in tissue culture. Greater numbers of macrophages could be isolated with further time in culture. After 72 h however, intrinsic glomerular cell contamination occurred. The majority of the cells were viable by fluorescein diacetate hydrolysis, and Trypan Blue exclusion. Further functional studies of these cells may provide some new insights into the cellular basis of macrophage-induced glomerular injury in experimental glomerulonephritis.

Mononuclear cell infiltrate and HLA-DR expression in low grade astrocytomas. An immunohistological study of 23 cases.

Frozen samples from 23 low grade (grade I and II) astrocytomas were studied by means of a panel of monoclonal antibodies to macrophages, lymphocytes (and their subsets) and HLA-DR antigens. Macrophages were present in low to moderate numbers in 38%-86% of cases, the variance in figures depending on the antibody used. T lymphocytes, the majority of CD8 phenotype, were detected in low numbers in 78% of tumours. B lymphocytes were scanty in 22% (5/22) and totally absent in the remaining cases. HLA-DR antigen was expressed by tumour cells in 35% (6/17) of cases. These findings indicate that in some low grade astrocytomas there is a mononuclear cell infiltrate with macrophages and secondarily CD8+ lymphocytes playing the major role. The significance of these findings remains speculative at present.

Regulation of 1,25-dihydroxyvitamin D3 production by cultured alveolar macrophages from normal human donors and from patients with pulmonary sarcoidosis.

Regulation of the production of the biologically active vitamin D3 sterol 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] by cultured pulmonary alveolar macrophages (PAM) obtained from 6 patients with pulmonary sarcoidosis and from 9 normal subjects was studied. The sarcoid cells, all collected from patients with normal calcium metabolism, synthesized 1,25-(OH)2-[3H]D3 from the substrate 25-hydroxyvitamin [3H]D3 (25OH-[3H]D3), whereas in vitro incubation with recombinant human interferon-gamma (IFN gamma) or lipopolysaccharide (LPS) was required for induction of synthesis of the hormone by normal PAM. Exogenous 1,25-(OH)2D3 (10-100 nmol/L) decreased endogenous hormone production by normal PAM by approximately 45%. The relative inhibitory effect of 1,25-(OH)2D3 was less pronounced in sarcoid PAM, in which 10-100 nmol/L 1,25-(OH)2D3 inhibited 250HD3-1-hydroxylase by approximately 25%. An accompanying induction of the 250HD3-24-hydroxylase, which is typical for renal cells, was found at low levels in only 3 of 10 experiments; in this regard, no differences between sarcoid and normal PAM were apparent. PTH or forskolin did not influence 250HD3 metabolism by PAM. 1,25-(OH)2D3 production by sarcoid PAM was enhanced by lipopolysaccharide and IFN gamma. Likewise, recombinant human interleukin-2 stimulated 1,25-(OH)2D3 production by sarcoid PAM, suggesting a possible role for both IFN gamma and interleukin-2 in the induction of 1,25-(OH)2D3 synthesis by sarcoid PAM in vivo. Recombinant human IFN alpha, IFN beta, and granulocyte-macrophage colony-stimulating factor had little effect. Dexamethasone and chloroquine, which have in vivo antihypercalcemic activity in sarcoidosis, both inhibited 1,25-(OH)2D3 synthesis by sarcoid PAM; chloroquine simultaneously stimulated the 24-hydroxylase. Our studies suggest that the 250HD3-metabolizing system in PAM is in some respects different from renal metabolism of 250HD3.

Cryptogenic abscess of the liver. Evidence of underlying reticuloendothelial cell dysfunction.

Evidence of underlying liver disease and reticuloendothelial system dysfunction was sought in five adults with histories of idiopathic or "cryptogenic" liver abscess. Although no evidence of underlying liver disease was obtained (median follow-up, 3.4 years; range, 2.1 to 4.8 years), four of five individuals demonstrated a marked impairment in their ability to clear antibody-tagged erythrocytes from the systemic circulation. The results of this study suggest that patients who develop cryptogenic abscesses of the liver have an underlying reticuloendothelial cell defect that may predispose them to liver abscess formation.

Changes of expression of peritoneal macrophages Fc and C3 receptors induced by transplantable melanomas.

Expression of Fc and C3 receptors was studied in the rosette tests on isolated peritoneal macrophages of control and melanoma-bearing hamsters. In hamsters with transplanted melanomas an increase of the percentage of macrophages with Fc and C3 receptor expression was observed. The increase was prominent among macrophages from animals with transplanted amelanotic melanoma, a tumor line with greater malignancy and changed antigenicity and immunogenicity.

Isolation and characterization of macrophages from scrapie-infected mouse brain.

We have isolated and characterized a population of brain macrophages from normal and scrapie-infected mice. The cells are phagocytic, possess Fc-IgG receptors, Mac-1 surface antigen and proliferate in the presence of macrophage colony stimulating factor. They resemble microglia in that they have a plasmalemmal distribution of the enzyme nucleoside diphosphatase, a property tht is characteristic of microglia in situ. In two of the three combinations of scrapie agent and mouse strain examined, the number of brain macrophages was several fold higher than in normal control mice. The increase was not observed in mice infected intraperitoneally or in control mice inoculated with normal brain homogenate. The increase is detectable as early as 3-5 weeks postinoculation. The agent/host combination that failed to show an increase in brain macrophages is one that develops large numbers of amyloid plaques. These observations suggest that these cells are closely associated with the scrapie pathogenic process in the CNS. The failure of these cells to increase in the plaque forming model of scrapie disease also suggests that they play a role in the control of CNS amyloidogenesis.

Total parenteral nutrition in mice bearing a metastatic carcinoma: tumor growth, metastasis and immunologic parameters.

The role of dietary manipulation of tumor growth, metastasis and immunologic parameters was studied in mice bearing Lewis lung carcinoma. Fourteen days following subcutaneous tumor implant, groups with tumor and their non-tumor bearing counterparts were assigned to one of the following feeding protocols: total parenteral nutrition (TPN), per oral (PO) intake of the parenteral diet, an oral casein diet (CAS), or electrolyte infusion plus the casein diet (ELECT). Intakes of energy and nitrogen were similar among all groups. Mice were killed 12 days later and peritoneal macrophages were tested for phagocytic activity. Tumor growth and metastasis were decreased from both infusion regimens with minimal loss of body weight as compared with casein fed mice. PO mice also showed lower tumor weight but metastasis was as great as in the casein group. Non-tumor-bearing infused mice showed depressed thymic weight, but thymic weight was not further reduced in tumor-bearing infused mice. PO feeding afforded no such protection in the presence of the carcinoma. Splenomegaly was observed in tumor-bearing mice on all regimens, but mice maintained on the parenteral diet demonstrated the largest proportion of macrophages containing nuclear debris. Analysis of free macrophages indicated no effect of diet regimen on non-immune phagocytic activity in both tumor-free and tumor-bearing mice. Possible alteration of splenic macrophage intracellular digestive capacity or phagocytic activity was suggested as a result of TPN.

Hematogenous origin of brain macrophages: a case report.

Del Río-Hortega believed that phagocytic cells in the CNS arise from microglia; however, recent autoradiographic studies have suggested a hematogenous origin for brain macrophages. To clarify this issue, we studied a young man with acute lymphoblastic leukemia and a peripheral white blood cell count of 22/mm3 who had an embolic brain infarct 2 weeks before he died. Macrophages were scarce within the lesion, suggesting that the principal phagocytic cell is hematogenous.

Stimulation of neovascularization by human rheumatoid synovial tissue macrophages.

Synovial tissue from patients with rheumatoid arthritis was enzymatically dissociated, and single cell suspensions were fractionated into subpopulations by centrifugation on continuous Percoll gradients. Five fractions (F1-F5) with densities of 0.991-0.998 gm/ml, 0.998-1.042 gm/ml, 1.042-1.062 gm/ml, 1.062-1.082 gm/ml, and 1.082-1.180 gm/ml, respectively, were prepared. F3 consistently contained the highest number of macrophages, while F2 and F4 contained substantially fewer macrophages. Macrophages present in F2, F3, and F4 were enriched by differential adherence to fibronectin-coated collagen gels. These macrophage-enriched cell preparations were found to be Fc and C3 positive, esterase positive, and peroxidase negative, to stain positively with anti-HLA-DR, anti-Leu-M3, OKM1, and OKM5 monoclonal antibodies, and to show characteristic features of macrophages by electron microscopy. Macrophages from F3 consistently induced neovascularization in rat corneas, while equal numbers of macrophages from F2 and F4 did not. Fibroblastic synovial cells and cells that did not adhere to fibronectin-coated collagen gels did not induce neovascularization. Within the rheumatoid synovium, there appears to be a major subpopulation of macrophages capable of inducing neovascularization, a process vital to the development of the rheumatoid synovial pannus.

Ectopic hematopoiesis in peritoneal tumor nodules induced by the myeloproliferative sarcoma virus in DBA/2 mice.

Tumor nodules composed of fibroblasts, large undifferentiated cells, granulocytes, and small lymphocytes develop in the spleens of adult DBA/2 mice infected with the myeloproliferative sarcoma virus (MPSV). They spread thereafter in the organism, and at the terminal stage of the disease they are especially numerous on the peritoneal membrane. The present study, performed on those tumor nodules to avoid contamination by exogenous hematopoietic cells, demonstrated that they were sites of granulopoiesis, which may have occurred via the local differentiation of granulomacrophage precursor cells (GM-CFC) and perhaps also from pluripotent hematopoietic stem cells, since these two populations were present in the tumor nodules (25 +/- 11 and 13 +/- 10, respectively, per 5-10(5) cells). Almost all (88%) those GM-CFC were able to clone in vitro without added colony-stimulating factor. A comparative study with the Moloney murine sarcoma virus-induced tumor indicated that the local production of hematopoiesis-stimulating factors was not sufficient to allow such ectopic granulopoiesis. These results imply the presence of a specific hematopoietic microenvironment in the MPSV-induced tumor nodules.

Studies of macrophage function in murine systemic lupus erythematosus. 3. The nature, anatomical location, and reversibility of the phagocytic defect.

The defect in phagocytosis and binding of antibody-coated sheep erythrocytes (EA) by peritoneal macrophages of (NZB X NZW)F1 or B/W mice is not intrinsic, but is related to the development of the autoimmune disease process. The defect appears to be confined to peritoneal macrophages, since bone marrow (BM)-derived macrophages have normal to elevated activities in vitro. The peritoneal macrophage defect is not due to blockade of Fc receptors in vivo, as shown by long-term culture or recovery of phagocytic and binding activities after removal of Fc receptors by pronase, but represents a reduced number of receptors with slightly delayed turnover. The defect can be reversed by elicitation of activated macrophages with Corynebacterium parvum, thioglycollate, or proteose peptone in vivo. Normal Fc-mediated phagocytosis and binding by BM-derived macrophages cultured from untreated autoimmune mice is enhanced by pretreatment of mice with C. parvum, thioglycollate, or proteose peptone. The cause of the defect in Fc-mediated phagocytosis by resident peritoneal macrophages of autoimmune mice was not ascertained; it may be due to abnormal macrophage kinetics or to the local effects of lymphokines released as a result of other autoimmune changes.