Hemolymph of triatomines presents fungistatic activity against Cryptococcus neoformans and improves macrophage function through MCP-I/TNF-α increase

Triatomines are blood-feeding arthropods belonging to the subfamily Triatominae (Hemiptera; Reduviidae), capable of producing immunomodulatory and water-soluble molecules in their hemolymph, such as antimicrobial peptides (AMPs). In this work, we evaluated the antifungal and immunomodulatory activity of the hemolymph of Meccus pallidipennis (MPH) and Rhodnius prolixus (RPH) against Cryptococcus neoformans. Methods: We assessed the activity of the hemolymph of both insects on fungal growth by a minimum inhibitory concentration (MIC) assay. Further, RAW 264.7 macrophages were cultivated with hemolymph and challenged with C. neoformans. Then, their phagocytic and killing activities were assessed. The cytokines MCP-1, IFN-γ, TNF-α, IL-10, IL-12, and IL-6 were measured in culture supernatants 4- and 48-hours post-infection. Results: Both hemolymph samples directly affected the growth rate of the fungus in a dose-dependent manner. Either MPH or RPH was capable of inhibiting fungal growth by at least 70%, using the lowest dilution (1:20). Treatment of RAW 264.7 macrophages with hemolymph of both insects was capable of increasing the production of MCP-I and TNF-α. In addition, when these cells were stimulated with hemolymph in the presence of C. neoformans, a 2- and a 4-fold increase in phagocytic rate was observed with MPH and RPH, respectively, when compared to untreated cells. For the macrophage killing activity, MPH decreased in approximately 30% the number of viable yeasts inside the cells compared to untreated control; however, treatment with RPH could not reduce the total number of viable yeasts. MPH was also capable of increasing MHC-II expression on macrophages. Regarding the cytokine production, MCP-I and TNF-α, were increased in the supernatant of macrophages treated with both hemolymphs, 4 and 48 hours after stimulation. Conclusion: These results suggested that hemolymph of triatomines may represent a source of molecules capable of presenting antifungal and immunomodulatory activity in macrophages during fungal infection.(AU)

Anti-obesity and LDL-cholesterol lowering effects of silkworm hemolymph in C57BL/6N mice fed high fat diet / 한국영양학회지

PURPOSE: Obesity, a worldwide epidemic, is associated with insulin resistance, hyperlipidemia, hypertension, cardiovascular disease, and certain cancers. Many strategies, including natural alternative anti-obesity agents, are used widely to prevent obesity. This study examined the effects of silkworm hemolymph on the weight control of C57BL/6N mice fed with a high-fat diet. METHODS: The mice were divided into five groups: normal group (N), high-fat diet group (HFC), high-fat diet and silkworm hemolymph (at dose of 1 mL/kg BW (HFS-1), 5 mL/kg BW (HFS-5) and 10 mL/kg (HFS-10) for 12 weeks. RESULTS: After 12 weeks treatment, the administration of silkworm hemolymph decreased the final body weight significantly along with a decrease in the weights of epididymal fat and total fat. The plasma LDL-cholesterol concentration was significantly lower in the HFS-1, HFS-5, and HFS-10 groups than in the HFC group. In addition, the leptin level of the HFS groups was significantly lower than those of the HFC group without a change in the plasma insulin concentration. CONCLUSION: These findings suggest that the silkworm hemolymph may have the potential to prevent obesity.

Estudio de componentes hemolinfáticos de Rhodnius prolixus y Triatoma maculata alimentados con diferentes fuentes sanguíneas / Study of hemolymphatic components of Rhodnius prolixus and Triatoma maculata fed on different blood sources

RESUMEN Introducción: Tradicionalmente, Rhodnius prolixus ha sido considerado el principal vector, mientras que Triatoma maculata resulta ser un vector secundario de Trypanosoma cruzi en el medio rural venezolano. En este trabajo se provee información de interés acerca de los componentes bioquímicos y del sistema inmune, humoral y celular, de la hemolinfa de R. prolixus y T. maculata alimentados sobre gallina y rata. Metodología: los insectos provenían de una colonia del laboratorio, mantenida a una temperatura de 27-29ºC, 50% de humedad relativa y ocho horas luz/día. Se analizó la hemolinfa de adultos alimentados sobre gallina y rata. Se observaron los hemocitos mediante microscopía óptica y electrónica de transmisión. Resultados: los parámetros bioquímicos de glucosa, lípidos y proteínas fueron afectados de forma distinta en R. prolixus y T. maculata según la fuente alimenticia. T. maculata presentó mayores niveles de actividad lítica de lisozima. Se observaron cuatro poblaciones de hemocitos (prohemocitos, plasmatocitos, granulocitos y oenocitos), cuyas características y medidas están acorde a lo reportado por otros autores sobre la subfamilia Triatominae. T. maculata presentó mayor número de prohemocitos y oenocitos que R. prolixus. Conclusiones: En su hemolinfa, R. prolixus y T. maculata son afectados distintamente en los componentes bioquímicos (glucosa, lípidos y proteínas) e inmunes, tanto humorales (lizosima) como celulares (prohemocitos, oenocitos), según sean alimentados sobre gallina y rata. Esto pone de manifiesto de la alimentación sobre uno de los componentes de la competencia vectorial, como lo es el sistema inmune propio de la hemolinfa de los insectos.

ABSTRACT Introduction: Traditionally, Rhodnius prolixus has been considered the main vector, while Triatoma maculata remains to be a secondary vector of Trypanosoma cruzi in the Venezuelan rural habitat. In this work, interesting information about the biochemical and immune (humoral and cellular) aspects of the hemolymph of R. prolixus and T. maculate, feeding on hen and rat, are presented. Methodology: Hemolymph was extracted from adult insects, maintained at 2729 °C with 50% of relative humidity and 8/16 (Light/Dark) hours, and fed on hen and rat. Hemocytes were observed by optical and transmission electron microscopy. Results: Biochemical parameters (glucose, lipids and proteins) varied in both species according to the food source. T. maculata presented higher levels of lysozyme lytic activity. Four hemocytes populations were observed by optical and transmission electron microscopy (prohemocytes, plasmocytes, granulocytes and oenocytes), exhibiting characteristics and measurements in accordance with previous literature related to the Triatominae subfamily. T maculata presented more prohemocytes and oenocytes than R. prolixus. Conclusions: On their hemolymph, R. prolixus and T. maculata are distinctly affected in their biochemical (glucose, lipids and proteins) and immune components, both humoral (lysozyme) and cellular (prohemocytes, oenocytes), depending on whether they were fed on hens or rats. Our results show that the food source affects the immune system of triatomines, and subsequently, their vectorial capacity may be compromised as well.

Allergenic Characterization of 27-kDa Glycoprotein, a Novel Heat Stable Allergen, from the Pupa of Silkworm, Bombyx mori

Boiled silkworm pupa is a traditional food in Asia, and patients with silkworm pupa food allergy are common in these regions. Still now only one allergen from silkworm, arginine kinase, has been identified. The purpose of this study was to identify novel food allergens in silkworm pupa by analyzing a protein extract after heat treatment. Heat treated extracts were examined by proteomic analysis. A 27-kDa glycoprotein was identified, expressed in Escherichia coli, and purified. IgE reactivity of the recombinant protein was investigated by ELISA. High molecular weight proteins (above 100 kDa) elicited increased IgE binding after heat treatment compared to that before heat treatment. The molecular identities of these proteins, however, could not be determined. IgE reactivity toward a 27-kDa glycoprotein was also increased after heating the protein extract. The recombinant protein was recognized by IgE antibodies from allergic subjects (33.3%). Glycation or aggregation of protein by heating may create new IgE binding epitopes. Heat stable allergens are shown to be important in silkworm allergy. Sensitization to the 27-kDa glycoprotein from silkworm may contribute to elevation of IgE to silkworm.

Evaluación de la actividad antibacterial de los extractos de cuerpos grasos y hemolinfa derivados de la mosca Sarconesiopsis magellanica (Diptera: Calliphoridae) / Assessment of antibacterial activity by fat bodies extracts and hemolymph derived from the blowfly Sarconesiopsis magellanica (Diptera: Calliphoridae)

Objetivo: Evaluar, en condiciones in vitro , la actividad antibacterial de los extractos de cuerpos grasos y de la hemolinfa de larvas de tercer estadio de Sarconesiopsis magellanica , la cual se comparó con los efectos obtenidos de las mismas sustancias derivadas de Lucilia sericata . S. magellanica (Diptera: Calliphoridae) es una mosca de importancia principalmente forense, utilizada en la determinación del intervalo post mortem . Por sus hábitos necrófagos, es considerada un modelo potencialmente útil en terapia larval. Material y métodos: Se extrajeron los cuerpos grasos de las larvas mediante la técnica de disección corporal y la hemolinfa se obtuvo mediante decapitación y centrifugación de los especímenes larvales. Las bacterias evaluadas fueron Staphylococcus aureus y Pseudomonas aeruginosa . Los métodos utilizados para evaluar la actividad antibacterial fueron difusión en agar y unidades formadoras de colonias (UFC/ml). Resultados: Después de la correspondiente incubación, los resultados generales mostraron que la actividad antibacterial de la hemolinfa y de los cuerpos grasos, tanto de L. sericata como de S. magellanica , fueron efectivos contra S. aureus y P. aeruginosa sin diferencias significativas entre las especies de moscas, aunque con algunas diferencias entre las cepas bacterianas. Conclusiones: Los resultados obtenidos sugieren que estas sustancias podrían tener un efecto similar en el tratamiento de heridas infectadas contra los microorganismos evaluados.

Objective: This study aimed to evaluate the in vitro antibacterial activity of fat body and hemolymph extracts from Sarconesiopsis magellanica (Diptera: Calliphoridae) third-instar larvae, compared to the effect obtained using the same extracts but derived from Lucilia sericata . S. magellanica blowflies are considered important in forensic sciences due to their usefulness in determining the post mortem interval. This blowfly could be useful in larval therapy due to its necrophagous habits. Materials and methods: Fat body from larvae was removed by dissection, and hemolymph via decapitation and centrifugation of larval specimens. The antibacterial effect was tested against Staphylococcus aureus and Pseudomonas aeruginosa using two methods: agar diffusion and colony forming units (CFU/mL). Results: Hemolymph and fat body extracts derived from both L. sericata and S. magellanica were effective against S. aureus and P. aeruginosa , with no significant differences between blowfly species, although with some differences between the bacterial strains. Conclusions: The results obtained suggest that S. magellanica and L. sericata fat body and hemolymph extracts might have a similar antimicrobial activity against these microorganisms when used to treat infected wounds.

Ovary histology and quantification of hemolymph proteins of Rhipicephalus (Boophilus) microplus treated with Melia azedarach / Histologia do ovário e quantificação de proteínas na hemolinfa de Rhipicephalus (Boophilus) microplus tratado com Melia azedarach

This study aimed to analyze ovary histology and quantify total protein in the hemolymph of Rhipicephalus (Boophilus) microplus females treated with hexane extracts from green fruits of Melia azedarach. Eight engorged females were immersed in the extract at 0.25% concentration, and eight in water containing 5% acetone (control). The females were dissected 72 hours after treatment, and the ovaries were weighed and subjected to standard histological techniques. The total protein concentration was measured in the hemolymph of 200 females, of which 100 were treated as described above and 100 served as a control. In the treated group, ovary weight reduction and predominance of immature oocytes were observed. In addition, there were decreases in the diameters of the cytoplasm and germ vesicle of the oocytes in the treated group, compared with the controls. The protein concentration in the hemolymph was higher in the treated group than in the controls. The morphological changes observed in the treated ovaries included: presence of vacuolization; alteration of oocyte morphology, which changed from rounded to elongated; deformation of the chorion; and disorganization of the yolk granules. These results demonstrate the action of M. azedarach fruit extracts on R. (B.) microplus oogenesis.

Este estudo foi desenvolvido, visando analisar a histologia do ovário e quantificar as proteínas totais na hemolinfa de fêmeas de Rhipicephalus (Boophilus) microplus tratadas com extrato hexânico de frutos verdes de Melia azedarach. Oito fêmeas ingurgitadas foram tratadas por imersão com o extrato na concentração de 0,25%, e oito com água contendo 5% de acetona (controle). As fêmeas foram dissecadas 72 horas após o tratamento e os ovários foram pesados e submetidos a técnicas histológicas padrões. A concentração total de proteína foi mensurada na hemolinfa de 200 fêmeas, sendo 100 tratadas como descrito anteriormente e 100 como controle. Foi observada redução do peso dos ovários, predomínio de ovócitos imaturos e houve diminuição nos diâmetros do citoplasma e da vesícula germinal dos ovócitos do grupo tratado em comparação ao controle. A concentração de proteína na hemolinfa foi mais alta no grupo tratado que no controle. As alterações morfológicas observadas nos ovários tratados foram a presença de vacuolizações, alteração da morfologia dos ovócitos que mudaram de redondos para alongados, deformação do córion e desorganização dos grânulos de vitelo. Estes resultados demonstram a ação do extrato de M. azedarach na ovogênese de R. (B.) microplus.

Induction of a gloverin-like antimicrobial polypeptide in the sugarcane borer Diatraea saccharalis challenged by septic injury

Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) is an important pest for Brazilian sugarcane. In the present study, we detected two distinct spots in hemolymph from septic injured larvae (HDs1 and HDs2), which are separated by 2DE gel electrophoresis. Both spots were subjected to in-gel tryptic digestion and MALDI-TOF/TOF analysis, which revealed the sequence VFGTLGSDDSGLFGK present in both HDs1 and HDs2. This sequence had homology and 80 percent identity with specific Lepidoptera antimicrobial peptides called gloverins. Analyses using the ImageMaster 2D software showed pI 8.94 of the HDs1 spot, which is similar to that described to Hyalophora gloveri gloverin (pI 8.5). Moreover, the 14-kDa molecular mass of the spot HDs1 is compatible to that of gloverins isolated from the hemolymph of Trichoplusia ni, Helicoverpa armigera and H. gloveri. Antimicrobial assays with partially purified fractions containing the HDs1 and HDs2 polypeptides demonstrated activity against Escherichia coli. This is the first report of antimicrobial polypeptides in D. saccharalis, and the identification of these peptides may help in the generation of new strategies to control this pest.

Antibodies raised against hemolymph of Anopheles culicifacies reduce the fecundity and malaria parasite development.

Background & objectives: Several studies have been made to study the effect of antisera raised against different tissues (hemolymh, ovary, midgut and salivary glands) on the fecundity and malaria parasite development in the different species of mosquitoes but there are no reports on the antisera raised against the hemolymph of Anopheles culicifacies, the principal malaria vector in India accounting for 65% of malaria cases. Hence, an attempt was made to study the same and evaluate its impact on malaria parasite development. Methods: Polyclonal and multifactorial antibodies were produced in rabbits against heterogenous mixture of hemolymph proteins. Antibodies against hemolymph proteins were screened for their potential to influence reproductive performance of mosquitoes. Antibody titer in rabbit serum was determined by ELISA and putative candidate antigens were identified in the hemolymph of An. culicifacies by western blotting. Cross reactivity amongst various tissues vis-a-vis hemolymph protein was also identified. In addition, a significant reduction in oocyst development was also observed in An. culicifacies mosquitoes that ingested antihemolymph antibodies along with Plasmodium vivax. Results: The maximum reduction in fecundity (57%) was observed during fourth week, after the last booster and number of oocyts per infected mosquito reduced by 73.35% in the group of mosquitoes that ingested antihemolymph antibodies along with the infected blood meal respectively. However, the ingestion of antibodies against hemolymph proteins did not have significant influence on hatchability. Antisera raised against hemolymph proteins of An. culicifacies recognized 11 polypeptides by western blotting. Interpretation & conclusion: During the present study, 11 putative candidate antigens were identified in the hemolymph of An. culicifacies, against which antibodies produced significantly reduced the fecundity by 57%. In addition, a significant reduction in oocyst development was also observed in An. culicifacies that ingested antihemolymph antibodies along with P.vivax.

Tripanosomatides like Trypanosoma theileri in the cattle tick Boophilus microplus / Tripanosomatídeos similares a Trypanosoma theileri no carrapato dos bovinos Boophilus microplus

Findings of epimastigotes forms of a tripanosomatide is reported in the hemolymph of the cattle tick Boophilus microplus in the state of Rio Grande do Sul, southern Brazil. Morphological evidences suggest they are similar to Trypanosoma theileri, a species described as non pathogenic to cattle, and usually transmitted by tabanids.

Descreve-se a ocorrência de formas epimastigotas de um tripanosomatideo na hemolinfa do carrapato do bovino Boophilus microplus no Estado do Rio Grande do Sul. Evidências morfológicas sugerem tratar-se de Trypanosoma theileri , espécie descrita como não patogênica aos bovinos e que usualmente é transmitida por tabanídeos.

Ocorrência de Borrelia spp. em cultura de células embrionárias do carrapato Boophilus microplus (Acari: Ixodidae) no Estado do Mato Grosso do Sul, Brasil / Occurrence of Borrelia spp. in culture of embryonic cells of the tick Boophilus microplus (Acari: Ixodidae) in the State of the Mato Grosso do Sul, Brazil

O presente trabalho teve como objetivo reportar a ocorrência de Borrelia spp. em culturas de células embrionárias de Boophilus microplus infectados naturalmente. Sete dias após o início de uma nova cultura primária de células embrionárias do carrapato B. microplus, incubadas a 31ºC, notou-se que as células começaram a degenerar. Ao exame em microscópio de contraste de fase detectou-se a presença de microrganismos alongado e com grande mobilidade. Lâminas de microscópio confeccionadas com amostras do sobrenadante da cultura, hemolinfa e massa de ovos, coradas pelo May Grünwald-Giemsa, permitiram a visualização de espiroquetas. O exame morfológico do microrganismo e sua visualização em B. microplus sugere ser Borrelia spp.

The aim of the present work was to report the occurrence of Borrelia spp. in embryonic cell cultures from naturally infected Boophilus microplus. Seven days after the beginning of a primary culture of embryonic cells of B. microplus at 31ºC was noted that the cells start suffering degeneration. Under examination at phase contrast microscope, the presence of prolongated microorganisms with great mobility was detected. Microscopic slides of the culture supernatant, hemolymph and egg mass, were stained by May Grünwald-Giemsa, allowing the visualization of the spirochetes. The morphologic examination of the microorganism and its visualization in. B. microplus, suggest to be Borrelia spp.

Lympho fibrinogen - a risk factor for coronary artery diseases

A study on 48 patients with coronary artery diseases at Internal Medicine Department in Hue Central Hospital from July to December/1998. Results: Patients who live in urban: 62.5% and rural: 37.5%. The risk factors seen in study group is tobacco smoking (50%), hypertension (50%) and increased cholesterol level (40%). The average plasma fibrinogen level in 48 cases in control group is 11.07± 1.90µmol/l. A relative risk of fibrinogen plasma for coronary artery diseases is 3.18 (confidential interval (CI) 95%). Attribute risk is 0.65%, it means 65% case coronary artery diseases due to high plasma fibrinogen level. The incidence risk of coronary artery diseases increase 9.06 times in people with increase plasma fibrinogen level. So the routine test on plasma fibrinogen has value in prevention and predict coronary artery diseases.

SDS-PAGE analysis of differentially expressed proteins from Anopheles stephensi hemolymph produced during the Plasmodium yoelii oocysts melanization / 第三军医大学学报

Objective To explore primarily the differentially expressed proteins in the hemolymph from adult female Anopheles stephensi ( An stephensi ) infected with Plasmodium yoelii ( P yoelii ) after being fed with sucrose solution containing nitroquine or not at different time points Methods Hemolymph of 2 groups of adult female An stephensi was collected with the expulsion method from the first day to the fifth day after the feeding Hemolymph samples were examined with SDS PAGE The protein gels were visualized by either Coomassie brilliant blue or silver staining, scanned and automatically analyzed by the BioRad1000 gel image analysis system for differential proteins bands Results On the second day of feeding with nitroquine, a few oocysts were partially melanized Furthermore, during the period from the fifth day to the ninth day, the number of mosquitoes with malanized oocysts and the number of melanized oocysts gradually increased The number of hemolymph protein binds in the treatment group was markedly more than that in the control Many different bands, mainly located at the molecular weight of (20～40)?10 3 and (60～80)?10 3, were visualized in the 2 groups The number of protein bands stained by the silver staining was more than that by the Coomassie brilliant blue staining Conclusion There are differentially expressed proteins in the hemolymph in An stephensi infected with P yoelii after being fed with sucrose solution containing nitroquine These differential proteins may be the melanization engaging proteins

Influence mechanisms of high temperature and low salinity on hemolymph basic proteins of Sinonovacula constricta / 中国海洋药物

Sinonovacula constricta(Lamarck), a kind of mollusks, is an important species of marine aquaculture. We choose it as our experiment sample and its basic protein expression profiles of hemolymph after short-term treatment ur(?)der high temperature and low salinity were detected, seven kinds of proteins or polypeptides were identified: Cysteine-rich protein 2 CRP2, Phospholipase A2, Pepidyl-prolyl cis- trans isomerase A, epidermal secretory protein xp2 precursor, brain-derived neurotrophic factor, tumor necrosis factor receptor, highmobility groupprotein HMGI-C. They have multiple functions, such as nutrition, gene expression regulation, protein synthesis, substances transport, antitoxin and anti-interference. The results suggested that the hemolymph played a key action in defense System of Sinonovacula constricta .

STUDY ON FREE AM1NO ACIDS AND PROTEIN IN HEMOLYMPH OF ANOPHELES STEPHEN SI / 中国寄生虫学与寄生虫病杂志

The changes in the contents of free amino acids in hemolymph of Anopheles stephenst were determined by automatic amino acid analyzer.The changes in hemolymph protein were determined by ultraviolet absorption method.Free amino acids in hemolymph of infected mosquitoes were compared with those in noninfected mosquitoes.At 4 days after blood meal,6 kinds of amino acids decreased markedly,and 5 kinds of amino acids increased markedly; at 7 days after blood meal,4 kinds of amino decreased markedly,while 7 kinds of amino accids increased markedly; at 11 days after cids blood meal,9 kinds of amino acids decreased markedly,and 4 kinds of amino acids increased remarkably.The protein concentration of infected mosquitoes was higher than that of noninfected ones.

Analysis of the protein in hemolymph from Anopheles stephensi during melanotic encapsulation of Plasmodium yoelii oocysts by two-dimensional electrophoresis / 第三军医大学学报

Objective To analyze the protein in hemolymph from adult female Anopheles stephensi (An. Stepheni) infected by Plasmodium yoelii after feeding with sucrose solution containing nitroquine or simple sucrose solution with two dimensional gel electrophoresis technique. Methods Hemolymphs from nitroquine fed, infected blood fed, and sucrose solution fed adult female An. stephensi were collected using the expulsion method on the third day after the feeding. Hemolymph protein concentration was examined with Bradford method. Then the hemolymph protein was analyzed by two dimensional electrophoresis. The protein spots were visualized by Coomassie brilliant blue staining. The spots were scanned and automatically analyzed by the ImageMaster VDS CL (Amersham Pharmacia) and ImageMaster 2D Elite software (Amersham Pharmacia). Results The protein concentration in the nitroquine fed group was always lower than that in the infected blood fed and sucrose solution fed groups. Two dimensional gel electrophoresis revealed 101 protein spots in nitroquine fed and 115 protein spots in the control with 51 matched, but unmatched 50 and 64 protein spots were detected in the treatment group and the control group, respectively. Different protein spots were mainly located at the molecular weight of (40-60)?10 3 and at the isoelectric points of basic end. Conclusion Two dimensional gel electrophoresis may directly reflect the difference of the protein. Both the difference of protein concentration and the protein spots may be involved in nitroquine induced melanotic encapsulation of Plasmodium yoelii oocysts.

LYMPHATIC PASSAGE AND DISTRIBUTION OF THE ERYTHROCYTE IN HEMOLYMPH NODE IN THE RAT / 解剖学报

The spatial structure of hemolymph node in the rat is studied by light, transmission and scanning electron microscopy after the fixation of arterial perfusion. The structure of hemolymph node is similar to that of the normal lymph node, and main characteristic is that a number of the red cells are seen in it. The erythrocytes were carried to the afferent lymphatic vessel and reach the medullary sinus, many erythrocytes travel through the rsubcapsula and cortical sinuses, and reach the lymphatic tissue of the local cortex with selectivity, and going through the paracortical zone and the sinus wall to the medulary sinus. Most of the red blood cells are phagocytosed by macrophages in the sinuses. The subcapsular and cortical sinuses of hemolymph nodes connect with medullary sinuses, and form a reticular lymphatic passage. Reticular cells in the sinuses constitute a spatial each other. There are macrophages, lymphocytes, plasmocytes and numerous red blood cells in the nets. Macrophages are anchored on the reticular cells by their pseudopodla, traping and phagocytosing the red cells and foreign matters. Sometimes a macrophage is found closely associated with lymphocytes.