PAMP induced expression of immune relevant genes in head kidney leukocytes of rainbow trout (Oncorhynchus mykiss)

Chettri, Jiwan Kumar; Raida, Martin; Holten-Andersen, Lars; Kania, Per W.; Buchmann, Kurt

doi:10.1016/j.dci.2010.12.001

Cited by 93 publications

(61 citation statements)

References 19 publications

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“…In addition to this particular gene regulation pattern found in b-glucan fed fish, it is interesting to note that the poly(I:C) had a greater effect on CRP and complement related gene up-or down-regulations across the three organs examined. This may suggest that MacroGard Ò stimulated CRP and complement responses to poly(I:C) injection and would support previous studies conducted on rainbow trout which have shown that poly(I:C) combined with zymosan or b-glucan significantly induced up-regulation of inflammatory cytokines such as IL-1b, TNF-a, IL-6 and IL-10 [55]. These results highlight the complexity of the carp immune acute phase response [36] and the fact that the determination serum acute phase protein profile may not be enough to estimate the degree of the immune response [38].…”

Section: Combined Action Of B-glucan and Pamps On Carp Immune Acute Psupporting

confidence: 77%

Feeding common carp Cyprinus carpio with β-glucan supplemented diet stimulates C-reactive protein and complement immune acute phase responses following PAMPs injection

Pionnier

Falcó

Miest

et al. 2014

Fish & Shellfish Immunology

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Section: Combined Action Of B-glucan and Pamps On Carp Immune Acute Psupporting

confidence: 77%

Feeding common carp Cyprinus carpio with β-glucan supplemented diet stimulates C-reactive protein and complement immune acute phase responses following PAMPs injection

Pionnier

Falcó

Miest

et al. 2014

Fish & Shellfish Immunology

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“…Our results confirm that increased transcriptional activity, previously demonstrated, that occurs in trout HKL in response to Zymosan A (Chettri et al, 2011), also causes increased secretion of pro-inflammatory cytokines TNF-α, IL-1β and IL-6. The result that best represents what has been described at the transcriptional level, is the secretion of IL-6, which at four hours of induction, achieved the highest detection in the culture medium, coinciding exactly with the previously demonstrated at transcriptional level, with the highest level of expression at the same time.…”

Section: Discussionsupporting

confidence: 80%

“…In the case of IL-1β and TNF-α, transcriptional activity described by Chettri et al (2011), increases to 12 h of incubation which, at protein level, does not correlate with our results, since at 10 h, basal levels of the cytokine in the culture medium are already detected. As to the concentration of inductor, it was proved that low doses of Zymosan A (0.1 mg mL -1 ) generate secretion of three cytokines, as well as it was shown previously occurring at transcriptional level.…”

Section: Discussioncontrasting

confidence: 45%

“…This OD at 450 nm phase is used for in vitro induction, both at 24 h of stabilization, as well as days later, where the population of cells adhering to the plate is in the 80% of the total cells grown at the beginning. HKL culture cells are capable of responding to the challenge using different PAMPs such as LPS, flagellin and Poly I:C, in the case of Zymosan A there has been an up regulation of pro-inflammatory genes like TNF-α, IL-1β, IL-6 and the key enzyme in the pathway of prostaglandins, COX-2 (Chettri et al, 2011). Our results confirm the ability of HKL to respond to the challenge with Zymosan A, in all the tested assays, both at enzyme level as in ELISAs.…”

Section: Discussionmentioning

confidence: 99%

“…This phase corresponds to a Percoll interface between 34% and 51%, which has been previously characterized in fish and particularly in rainbow trout (Chettri et al, 2010(Chettri et al, , 2011. This OD at 450 nm phase is used for in vitro induction, both at 24 h of stabilization, as well as days later, where the population of cells adhering to the plate is in the 80% of the total cells grown at the beginning.…”

Section: Discussionmentioning

confidence: 99%

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Parameters to evaluate the immunostimulant effect of Zymosan A in head kidney leucocytes (HKL) of salmonids

Abarca

Bethke

Narváez

et al. 2012

lajar

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ABSTRACT. The interest in characterizing fish immune responses against pathogens or discovering new immunostimulant molecules requires the use of mass screening techniques. The classical model has been the challenge in 1000 L tanks, using a high number of fishes, resulting in high economic cost. In this work, the purpose was to minimize the costs of such tests using in vitro cellular models. In primary cultures of head kidney leucocytes (HKL) immune response to Zymosan A, a type of β-glucan, was evaluated. The quantification and detection of pro-inflammatory cytokines was done based on both enzymatic activity and ELISA. The activity of phagocyte oxidase and inducible nitric oxide synthase (iNOS) was measured indirectly through NBT reduction by reactive oxygen species and Griess reaction to nitric oxide, respectively. For detecting pro-inflammatory cytokines such as TNF-α, IL-6 and IL-1β, monospecific antibodies generated in our laboratory were used by indirect ELISA. The immune parameters were evaluated at different concentration of Zymosan A and the evolution of the immune response over time was established. The results suggest the better time for evaluating the immunostimulant capacity of glucans and the appropriate dose of it, which is required to demonstrate the activation of macrophages (ROS and iNOS) and the production of proinflammatory molecules, both as indicators of good immune response. Currently, work is being conducted improving these methods, creating an ELISA sandwich and generating antibodies against total proteins of HKL for pre-absorption of induced samples. In particular, the implementation of ELISA for the characterization of cytokine secretion profiles is a biotechnological contribution in aquaculture, which has not been implemented before this work. Keywords: head kidney leucocytes, Zymosan A, fish innate immunity. salmonids, Chile.Parámetros para la evaluación del efecto de Zimosán A como inmunoestimulante sobre leucocitos de riñón cefálico (HKL) de salmónidos RESUMEN. El interés en la caracterización de la respuesta inmune de peces contra patógenos o el descubrimiento de nuevas moléculas inmunoestimulantes, requiere el uso de técnicas de uso masivo y rápido. El modelo clásico ha sido el desafío en estanques de 1000 L, con un elevado número de peces, lo que tiene un alto costo económico. El objetivo de este trabajo fue reducir al mínimo los costos de dichos ensayos, utilizando modelos celulares in vitro. Para ello, se evaluó el efecto del inmunoestimulante de Zymosan A, un tipo de β-glucano, sobre la respuesta inmune en cultivos primarios de leucocitos de riñón cefálico (HKL), mediante la cuantificación enzimática y la detección de citoquinas pro-inflamatorias. La actividad de la fagocito oxidasa y de la óxido nítrico sintetasa inducible (iNOS), se midió indirectamente a través de la reducción de NBT, por especies reactivas de oxígeno, y la reacción de Griess para óxido nítrico, respectivamente. La detección de citoquinas pro-inflamatorias como el TNF-α, IL-6 e IL-1β, se realizó mediante E...

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Black Scorpionfish (Scorpaena porcus) Hemopoiesis: Analysis by Flow Cytometry and Light Microscopy

Andreyeva

Солдатов

Кухарева

2017

The Anatomical Record

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Cell suspensions of head kidney and spleen of black scorpionfish (Scorpaena porcus L.) have been studied using flow cytometry and light microscopy. On the basis of forward scatter (FS) and side scatter (SS) distribution and light microscopy, two main types of cells in the hemopoietic organs were identified: "small cells" (5.1-8.9 mm) and "large cells" (10.8-15.3 mm). Subpopulation of small cells was formed by thrombocytes, lymphocytes, and elements with functionally inactive nuclei. Euchromatin level in their nuclei was low and acidophilic cytoplasm corresponded to negligible nucleic acids content. No proliferative activity was observed using SYBR Green I fluorescence analysis. Morphological characteristics of these cells coincided with colony forming units of mammals. Large cells in head kidney consisted of two subpopulations of cells differing in granularity and DNA content. Proliferating blast cells and differentiating cells of all hemopoietic lines were identified among them. Macrophages and apoptotic cells were also detected in head kidney. In spleen large cells cluster mainly included aged red blood cells with extended lengthwise axis. Blast cells and differentiating elements in spleen were not observed.

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PAMP induced expression of immune relevant genes in head kidney leukocytes of rainbow trout (Oncorhynchus mykiss)

Cited by 93 publications

References 19 publications

Feeding common carp Cyprinus carpio with β-glucan supplemented diet stimulates C-reactive protein and complement immune acute phase responses following PAMPs injection

Feeding common carp Cyprinus carpio with β-glucan supplemented diet stimulates C-reactive protein and complement immune acute phase responses following PAMPs injection

Parameters to evaluate the immunostimulant effect of Zymosan A in head kidney leucocytes (HKL) of salmonids

Black Scorpionfish (Scorpaena porcus) Hemopoiesis: Analysis by Flow Cytometry and Light Microscopy

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