Effect of vaginally administered (Ala8,13,18)-magainin II amide on the morphology of implantation stage endometrium in the rhesus monkey (Macaca mulatta)

Ghosh, Debabrata; Dhawan, Latika; Lalitkumar, P. G. L.; Wong, Viviana; Rosario, J.F.; Hendrickx, Andrew G.; Lasley, Bill L.; Overstreet, James W.; Sengupta, Jayasree

doi:10.1016/s0010-7824(01)00211-6

Cited by 3 publications

(3 citation statements)

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“…The growth factors like FGF2 and CSF1 which are known to support the growth and differentiation of placental trophoblasts [ 38 – 40 ] were also down-regulated by AMA. Collectively, our results provide probable physiological basis of our earlier observation that application of AMA resulted in the breakdown of cellular homeostasis in isolated first trimester villous cytotrophoblasts [ 9 , 10 ].…”

Section: Discussionsupporting

confidence: 74%

Cationic antimicrobial peptide, magainin down-regulates secretion of pro-inflammatory cytokines by early placental cytotrophoblasts

Anupa

Bhat

Srivastava

et al. 2015

Reprod Biol Endocrinol

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BackgroundHuman placental villous cytotrophoblasts exhibit relative externalization of negatively charged moieties to the outer leaflet of the plasma membrane during the time of syncytialization rendering their reactivity to positively charged cationic antimicrobial peptides (CAMPs) during the window of implantation and early placentation. Vaginal administration of a synthetic CAMP, Ala8,13,18-magainin II amide (AMA) inhibited blastocyst implantation and early placentation in monkeys. Furthermore, the administration of AMA resulted in significant inhibition of cell differentiation, enhancement in apoptosis and loss of viability in first trimester placental villous cytotrophoblasts in primary culture. The present study examines the effect of in vitro application of different doses (0, 1, 10, 100, 1000 ng/ml) of AMA on the secreted cytokine profiles of cytotrophoblasts obtained from placental villi samples (n = 13) collected during 8–9 weeks of gestation and grown on three-dimensional collagen matrix in vitro.MethodsA panel of forty-eight (48) cytokines in conditioned medium was analysed using multiplex immunoassays technique. Further, the steady state transcript levels of four cytokines (CCL4, CCL5, IL1B, IL6), the concentrations of which were affected by AMA in the isolated cytotrophoblasts, as well as, two cytokines (IL1A and TNF) which were not affected by AMA were estimated. Input list of cytokines secreted by cytotrophoblasts and showing differential secretion in response to AMA were used in enrichment analysis for the generation of biological networks.ResultsPlacental cytotrophoblasts secreted 27 cytokines, 13 of which are affected by AMA in vitro with significantly decreased secretion of CCLs-2, 3, 4, 5, CXCLs-1 and 8, FGF2 and MCSF and that of IL1B, IL6 and MIF, and increased secretion of IL16 and IL-2RA. Of the above cytokines showing differential secretion, only IL-2RA, IL16 and MIF showed significant correspondence in the steady state expression of their respective transcript levels. Post-hoc Enrichment analysis revealed Toll-like receptor (TLR) mediated pathways were the top-scored target pathways that were affected by AMA.ConclusionsAdministration of a CAMP causes shift in the balance of immune-inflammatory responses involving downstream pathways of TLRs in cytotrophoblast function. Further verification of functions of placental trophoblasts on administration of CAMP with pregnancy outcome is necessary.Electronic supplementary materialThe online version of this article (doi:10.1186/s12958-015-0119-8) contains supplementary material, which is available to authorized users.

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Section: Discussionsupporting

confidence: 74%

Cationic antimicrobial peptide, magainin down-regulates secretion of pro-inflammatory cytokines by early placental cytotrophoblasts

Anupa

Bhat

Srivastava

et al. 2015

Reprod Biol Endocrinol

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“…These actions of AMA collectively might have resulted in observed pregnancy inhibition in monkeys [9,15]. …”

Section: Discussionmentioning

confidence: 99%

“…In a previous study, we have reported that intra-vaginal administration of AMA resulted in the inhibition of blastocyst implantation in the rhesus monkey [9]. Further study in the same species indicated that the anti-nidatory effect of AMA might be resulted from its inhibitory effect on trophoblast cell differentiation [15]. However, little is known about the cellular behavior of early placental trophoblast cells in the presence of cationic antimicrobial peptides.…”

Section: Introductionmentioning

confidence: 99%

In-vitro effects of the antimicrobial peptide Ala8,13,18-magainin II amide on isolated human first trimester villous trophoblast cells

Sengupta

Khan

Huppertz

et al. 2011

Reprod Biol Endocrinol

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BackgroundResearch on antimicrobial cationic peptides (AMPs) has gained pace toward using their potential to replace conventional antibiotics. These peptides preferentially interact with negatively charged membrane lipids typically seen in bacteria and thereby lead to membrane perturbations and membrane dysfunction. However, one possible disadvantage of AMP drugs is their potential for toxicity, especially to those cells which display externalization of negatively charged moieties to the outer leaflet of the plasma membrane during the process of syncytialization. Human placental villous trophoblast is one such cell type. Indeed, intra-vaginal administration of an antimicrobial cationic peptide Ala8,13,18-magainin II amide (AMA) which is a synthetic analogue of magainin 2 derived from Xenopus frog has been observed to result in inhibition of pregnancy establishment in monkeys. However, only little is known about the cellular behavior of early placental cytotrophoblasts (CTB) in the presence of cationic antimicrobial peptides. It is believed that suitable cell culture approaches using AMA as a representative alpha-helical AMP may yield tangible knowledge in this regard.MethodsImmunocytochemical (ICC) analyses using confocal microscopy (n = 6 for each treatment sub-group) and Western blot (WB) method (n = 5 for each treatment sub-group) of CTB differentiation based on synthesis of beta-hCG and hPL, and apoptosis based on apoptosis-associated cytokeratin 18 neo-epitope (CK18f) were performed for CTB isolated from human first trimester placental villi and grown in serum-free primary culture for 24 h, 48 h and 96 h on rat-tail collagen with and without AMA (1000 ng/ml). Moreover, secretion of beta-hCG and hPL into conditioned media from isolated CTB grown in vitro for 24 h, 48 h and 96 h (n = 6/each sub-group) with and without AMA was examined using enzyme immunoassays. Furthermore, TUNEL assay, and cell viability based on LDH leakage into medium (n = 6/each sub-group) were assessed to examine the phenomenon of cell death with time and administration of AMA.ResultsCTB in serum-free primary culture showed increased (P < 0.05) level of synthesis and secretion of beta-hCG and hPL with time, and higher (P < 0.05) level of cellular cytokeratin 18 neo-epitope and number of TUNEL-positive cells, and LDH activity in conditioned medium at 96 h of culture. Exposure of CTB to AMA resulted in lower (P < 0.05) level of synthesis and secretion of beta-hCG and hPL, as well as, an increase (P < 0.05) of cellular cytokeratin 18 neo-epitope and number of TUNEL-positive cells, and LDH activity in conditioned medium at 96 h as compared to the control treatment.ConclusionsAdministration of AMA resulted in attenuation of differentiation, enhancement in apoptosis and loss of viability in early placental villi trophoblast cells in primary culture. Thus, it appears that administration of alpha-helical AMP may adversely affect the process of placentation and pregnancy outcome.

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Magainins – A Model for Development of Eukaryotic Antimicrobial Peptides (AMPs)

Dennison

Harris

Phoenix

2014

Novel Antimicrobial Agents and Strategies

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Effect of vaginally administered (Ala8,13,18)-magainin II amide on the morphology of implantation stage endometrium in the rhesus monkey (Macaca mulatta)

Cited by 3 publications

References 14 publications

Cationic antimicrobial peptide, magainin down-regulates secretion of pro-inflammatory cytokines by early placental cytotrophoblasts

Cationic antimicrobial peptide, magainin down-regulates secretion of pro-inflammatory cytokines by early placental cytotrophoblasts

In-vitro effects of the antimicrobial peptide Ala8,13,18-magainin II amide on isolated human first trimester villous trophoblast cells

Magainins – A Model for Development of Eukaryotic Antimicrobial Peptides (AMPs)

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