In vitro activation of fish phagocytic cells by GH, prolactin and somatolactin

1996 ◽  
Vol 151 (1) ◽  
pp. 113-118 ◽  
Author(s):  
M Sakai ◽  
M Kobayashi ◽  
H Kawauchi

Abstract The activation of rainbow trout (Oncorhynchus mykiss) phagocytic cells by chum salmon GH, prolactin (PRL) and somatolactin (SL) was investigated in vitro. Rainbow trout kidney leucocytes were cultured in RPMI 1640 medium containing 1, 10, 50 or 100 ng of each hormone/ml and the production of superoxide anion was measured using reduction of nitroblue tetrazolium (NBT) and ferricytochrome C. Macrophages incubated with 10–100 ng GH or PRL/ml showed significantly enhanced production of superoxide anion in both the NBT and ferricytochrome C tests compared with control macrophages (without hormone). SL did not induce enhanced production of superoxide anion in macrophages. The phagocytic cells treated with GH or PRL also showed increased phagocytic activity and phagocytic index. However, the cells treated with SL showed no enhancement of phagocytic activity or index. These results indicate that GH and PRL stimulate macrophages in vitro. Journal of Endocrinology (1996) 151, 113–118

2014 ◽  
Vol 17 (3) ◽  
pp. 453-458 ◽  
Author(s):  
J. Małaczewska ◽  
A. K. Siwicki ◽  
R. Wójcik ◽  
W. a. Turski ◽  
E. Kaczorek

Abstract Kynurenic acid (KYNA), an endogenous neuroprotectant formed along the kynurenine pathway of tryptophan degradation, is a selective ligand of the GPR35 receptor, which can be found on the surface of various populations of human immune cells. In infections and inflammations, KYNA produces an anti-inflammatory effect through this receptor, by depressing the synthesis of reactive oxygen species and pro-inflammatory cytokines. However, it is still unrecognized whether receptors for kynurenic acid are also localized on immune cells of poikilothermic animals, or whether KYNA is able to affect these cells. The objective of this study has been to determine the effect of different concentrations of kynurenic acid (12.5 μM to 10 mM) on the viability and mitogenic response of lymphocytes and on the activity of phagocytic cells isolated from blood and the spleen of rainbow trout. The results imply low toxicity of kynurenic acid towards fish immune cells, and the proliferative effect observed at the two lowest concentrations of KYNA (12.5 μM and 25 μM) seems indicative of endogenous kynurenic acid being capable of activating fish lymphocytes. Non-toxic, micromole concentrations of KYNA, however, had no influence on the mitogenic response of lymphocytes nor on the activity of phagocytes in rainbow trout under in vitro conditions. There is some likelihood that such an effect could be observed at lower, nanomole concentrations of KYNA.


Author(s):  
Triana Hertiani ◽  
Agustinus Yuswanto ◽  
Sylvia Utami Tunjung Pratiwi ◽  
Harlyanti Mashar

Massoia (Massoia aromatica Becc., Lauraceae) bark has been widely used as a component of traditional Indonesian medicine. The indigenous people boil or steam the bark for traditional applications. Our preliminary research revealed the potency of Massoia essential oil and its major compound, C-10 Massoialactone as potential immunomodulator in vitro. However, no scientific evidence regarding its in vivo effects is available. Therefore, this study evaluated the potential immunomodulatory effects of Massoia bark infusion on the nonspecific immune response (phagocytosis) of Wistar rats. The aqueous extract of Massoia bark was obtained by boiling pulverized bark in water, and the C-10 massoialactone content of the extract was determined through Thin Layer Chromatography (TLC) densitometry. For the in vitro assay, macrophages were treated with the freeze-dried infusion at the concentrations of 2.5, 5, 10, 20, or 40 μg/mL media. For the in vivo assay, 2-month-old male Wistar rats were divided into 5 groups. The baseline group received distilled water at the dose of 1 mL/100 g BW with the immunostimulant herbal product “X” administered as the positive control at the dose of 0.54 mL/rat. The treatment groups received the infusion at a dose of 100, 300, or 500 mg/100 g BW. Treatments were given orally every day for 14 days. The ability of macrophage cells to phagocyte latex was determined as phagocytic index (PI) and was observed under microscopy with 300 macrophages. The in vitro study revealed that the phagocytic activity of the infusion-treated macrophages significantly increased in comparison with that of the control macrophages in a concentration-dependent manner. Among all treatment concentrations, the concentration of 40 μg/ml provided the highest activity with a PI value of 70.51% ± 1.11%. The results of the in vivo assay confirmed those of the in vitro assay. The results of the present study indicate that Massoia bark can increase the phagocytic activity of rat macrophage cells. Its potential as a naturally derived immunomodulatory agent requires further study.


2000 ◽  
Vol 46 (3) ◽  
pp. 25-28
Author(s):  
B. A. Bakhmetev ◽  
N. S. Likhacheva

Effects of somatotropic hormone in different concentrations on the phagocytic activity of various populations of human lymphocytes and production of superoxide anion were analyzed in vitro. Blood cells were incubated in medium 199 with various hormone concentrations for 1 h at 37°C. Besides the traditional general parameters of phagocytosis, the counts of eosinophils, neutrophils, monocytes phagocytizing and not phagocytizing sheep red cells (SRC) were evaluated with regard to their activity (number of phagocytosed SRC) in the total pool of phagocytes (sum of all phagocytizing cells of different activity per mm3 blood). The spontaneous and zymosan-stimulated levels of superoxide anion were evaluated in the nitroblue tetrasolium reduction test. Addition of somatotropic hormone stimulated the total phagocytic activity of human leukocytes by activating al! types of phagocytes, but the hormone effect was different for different cells. Growth hormone exerted the greatest stimulating effect on monocytes, increasing 3-5-fold their role in total phagocytosis, in comparison with the control. Addition of growth hormone increased zymosan-stimulated production of superoxide anion but did not change its spontaneous level. The results indicate the probability and demonstrate the direction of the immediate effect of growth hormone on the functions of some populations of human peripheral blood leukocytes.


1991 ◽  
Vol 260 (5) ◽  
pp. R969-R976
Author(s):  
A. P. Bautista ◽  
N. B. D'Souza ◽  
C. H. Lang ◽  
J. Bagwell ◽  
J. J. Spitzer

Bacterial endotoxins [lipopolysaccharide (LPS)] are potent immunomodulators, and ethanol is known to depress certain immune defense mechanisms. Thus the combined impact of these two agents on the generation of superoxide anion (O2(-).) by isolated hepatic phagocytic cells was investigated. Ethanol was infused intravenously into rats for 7 h, and Escherichia coli LPS was injected intravenously at 4 h after ethanol administration. Control groups received an equal volume of saline or ethanol alone. Nonparenchymal cells that were composed of endothelial and Kupffer cells and few polymorphonuclear neutrophils (PMN; less than 1%) were obtained after collagenase-pronase digestion. In the LPS-treated rats, the total number of PMN per liver increased significantly. Histological sections of the liver showed PMN infiltration and areas of necrosis after LPS treatment with or without ethanol. In the presence of either phorbol 12-myristate 13-acetate or opsonized zymosan in vitro, Kupffer cells and hepatic PMN from LPS-treated rats generated large amounts of O2(-).. Ethanol intoxication in vitro by these cells to 50%. Ethanol alone (without LPS) had no effect on the production of O2(-).. These studies demonstrate that ethanol intoxication was associated with the downregulation of the LPS-enhanced in vivo priming of hepatic phagocytes to generate O2(-). in vitro and may thus contribute to the enhanced susceptibility of alcoholic subjects to develop an infection.


10.5219/1238 ◽  
2020 ◽  
Vol 14 ◽  
pp. 328-335
Author(s):  
Muhamad Fauzi Ramadhan ◽  
Nurkhasanah Mahfudh ◽  
Nanik Sulistyani

Immunomodulators are pharmacological agents that modify or regulate the immune system through stimulating the functioning of the immune system and, at the same time, inhibiting excessive immune responses. This study was conducted to determine the active compound in Z. cassumunar that is responsible for increasing the immune system based on the parameters of phagocytic activity. The isolation method began with fractionation, which involved extraction with ethanol and successive fractionation with hexane and chloroform. Z. cassumunar extract, hexane fraction, and chloroform fraction were tested on mice macrophage cells for their phagocytic functions. The phagocytic activity of macrophages was measured by active phagocytic cells (averagely 39.194 ±1.597, 27.923 ±2.941, and 62.090 ±6.947) and phagocytic index (in a row, averagely 47.513 ±2.844, 41.129 ±7.195, and 101.527 ±10.555). The results showed that the Z. cassumunar extract,  hexane fraction, and chloroform fraction exhibited more significant phagocytic activities of macrophages (p <0.05) compared with the normal group. Since the chloroform fraction showed the best result, this fraction was further separated by column chromatography. This procedure yielded five sub-fractions, namely F1, F2, F2C, F3, and F4. Based on the phagocytic activity testing, the results were as follows: (1) the active phagocytic cells of F1, F2, F2C, F3 and F4 were 18.860 ±3.191, 27.077 ±4.482, 15.749 ±3.026, 64.333 ±1.780, and 44.943 ±2.944, respectively, and (2) the phagocytic indices were 30.0249 ±3.4231, 44.5969 ±8.3646, 24.5597 ±5.4487, 102.7447 ±1.0806, and 76.5007 ±4.7293. Because F3 produced the best result, this subfraction was then identified using 1H-NMR and 13C-NMR. The identification results showed that F3 was (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-ol as an active compound.


1993 ◽  
Vol 39 (11) ◽  
pp. 1051-1058 ◽  
Author(s):  
Rafael A. Garduño ◽  
Julian C. Thornton ◽  
William W. Kay

A model was developed to study the fate of the fish pathogen Aeromonas salmonicida in vivo, inside a specialized intraperitoneal chamber implanted in rainbow trout, Oncorhynchus mykiss. Although normally recalcitrant to lytic agents in vitro, owing to the presence of its regular surface array (S layer), A. salmonicida was rapidly killed in the peritoneal cavity by a host-derived, soluble lytic activity present in peritoneal fluid. Peritoneal fluid was also found to kill other bacteria and lyse various types of erythrocytes, but was particularly lytic to A. salmonicida. Intraperitoneal survival of injected (free) A. salmonicida cells was several orders of magnitude higher than survival of implanted (restrained) cells. Injected free cells could evade the lytic activity of peritoneal fluid because they readily spread, initiating lethal infections. One evasion strategy was envisioned to be the penetration of peritoneal and (or) tissue macrophages. In spite of the killing mechanisms of these phagocytic cells, A. salmonicida was still able to survive and even replicate inside head kidney macrophages, thereby supporting the notion of A. salmonicida as a facultatively intracellular pathogen. Intraperitoneal chambers in rainbow trout may constitute a valuable experimental tool for studying the in vivo fate of A. salmonicida, and perhaps of other fish pathogens as well.Key words: Aeromonas salmonicida, intraperitoneal chambers, rainbow trout, complement-mediated cell lysis.


2018 ◽  
Vol 17 (2) ◽  
pp. 161-165
Author(s):  
Sri Wahdaningsih ◽  
Subagus Wahyuono ◽  
Sugeng Riyanto ◽  
Retno Murwanti

The peel of red dragon fruit [Hylocereus polyrhizus (F.A.C.Weber) Britton and Rose] can be used to treat various diseases and to improve immune system of body. This study was aimed to investigate the in vitro macrophage phagocytic activity of extracts and fractions of red dragon fruit peel (Hylocereus polyrhizus). The in vitro test was conducted based on the method of Leijh et al. The parameters of phagocytic activity were based on the macrophages capacity to phagocytose latex beads using the calculation of phagocytic capacity and phagocytic index. The test results indicated significant difference (p < 0.05). The petroleum benzene extract showed higher phagocytic activity of macrophages than methanol extract of the fruit peel, sediment, and media control (-). The LSD test showed that macrophage phagocytic activity using fractions (500 and 100 μg/ml) was significantly different from macrophage phagocytic activity using fractions (20 μg/ml), sediment (500, 100 and 20 μg/ml), extracts (500, 100 and 20 μg/ml), and media control. Dhaka Univ. J. Pharm. Sci. 17(2): 161-165, 2018 (December)


2012 ◽  
Vol 9 (4) ◽  
pp. 651-655
Author(s):  
Baghdad Science Journal

The aqueous extract of Citrullius colocynthis dried seeds (160 ?g/ml) was in vitro evaluated for its effect on phagocytic index (PI) and lymphocyte transformation index (LTI) of blood cells obtained from 30 apparently healthy blood donors (15 males and 15 females). The PI was further in vivo evaluated in cells of peritone, spleen and liver of mice treated with the extract at a dose of 0.64 mg/kg. The results revealed that in in vitro study, phagocytic cells treated with the extract showed a significant increased percentage as compared with untreated cells (60.0 vs. 44.1%). Phagocytes obtained from peritone (44.1 vs. 30.0%) and spleen (45.6 vs. 39.6 %) of treated and untreated mice behaved in a similar manner, while liver phagocytes showed no significant difference between PI of immunological function of the investigated cells, and may use as therapeutic agent. treated and untreated mice. For LTI, cultures I and II shared an approximated mean (70.0 and 68.0%, respectively), but both indices were significantly higher than the recorded LTI in culture III (54.0%). These findings suggest that the plant extract is effective in enhancing the


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