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Placenta2012; 33(5); 379-386; doi: 10.1016/j.placenta.2012.01.018

Molecular evidence for natural killer-like cells in equine endometrial cups.

Abstract: To identify equine orthologs of major NK cell marker genes and utilize them to determine whether NK cells are present among the dense infiltration of lymphocytes that surround the endometrial cup structures of the horse placenta during early pregnancy. Methods: PCR primers were developed to detect the equine orthologs of NKP46, CD16, CD56, and CD94; gene expression was detected in RNA isolated from lymphocytes using standard 2-step reverse transcriptase (RT) PCR and products were cloned and sequenced. Absolute real-time RT-PCR was used to quantitate gene expression in total, CD3+, and CD3- peripheral lymphocytes, and invasive trophoblast. Lymphocytes surrounding the endometrial cups (ECL) of five mares in early pregnancy were isolated and NK marker gene expression levels were assayed by quantitative RT-PCR. Methods: Absolute mRNA transcript numbers were determined by performing quantitative RT-PCR and comparing values to plasmid standards of known quantities. Results: NKP46 gene expression in peripheral CD3- lymphocytes was higher than in CD3+ lymphocytes, CD16 levels were higher in the CD3+ population, and no significant differences were detected for CD56 and CD94 between the two groups. Expression of all four NK cell markers was significantly higher in lymphocytes isolated from the endometrial cups of pregnant mares compared to PBMC isolated from the same animal on the same day (NKP46, 14-fold higher; CD94, 8-fold higher; CD16, 20-fold higher; CD56, 44-fold higher). Conclusions: These data provide the first evidence for the expression of major NK cell markers by horse cells and an enrichment of NK-like cells in the equine endometrium during pregnancy.
Copyright © 2012 Elsevier Ltd. All rights reserved.
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Publication Date: 2012-02-21 PubMed ID: 22357194PubMed Central: PMC3319276DOI: 10.1016/j.placenta.2012.01.018Google Scholar: Lookup
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  • Journal Article
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  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research investigates the presence of natural killer (NK) cells in the horse’s placenta during early pregnancy, by identifying horse versions of key NK cell marker genes. The researchers found high expressions of these markers, suggesting a prevalence of NK-like cells in the endometrium during pregnancy.

Research Methods

  • To detect the presence of NK cells, PCR primers were utilized to detect horse orthologs of NK cell marker genes (NKP46, CD16, CD56, and CD94). These genes are commonly used to identify NK cells.
  • The gene expression was recognized in the RNA isolated from lymphocytes, the white blood cells that play a significant role in immune response, using the standard 2-step reverse transcriptase PCR technique.
  • Once the genes were detected, they were cloned and sequenced to ensure accuracy.
  • Following this, an absolute real-time PCR technique was used to quantify the gene expression in lymphocytes and invasive trophoblast – cells that provide nutrients to the embryo and develop into a large part of the placenta.
  • Lymphocytes around the horse’s endometrial cups (ECL) during early pregnancy were isolated and their NK marker gene expression was assayed utilizing quantitative PCR.

Results Analysis

  • The expression levels of the NK cell markers in both lymphocytes, CD3+ and CD3- types, were analyzed. CD3- lymphocytes revealed higher NKP46 gene expression compared to CD3+ lymphocytes while CD16 levels were higher in CD3+ lymphocytes.
  • When the researchers compared cell markers in lymphocytes from the endometrial cups of pregnant horses with peripheral blood mononuclear cells (PBMC) from the same horses, they found significantly higher marker expression in the former (NKP46, 14-fold higher; CD94, 8-fold higher; CD16, 20-fold higher; CD56, 44-fold higher). This data suggests the existence of major NK cell markers in horse cells and a higher prevalence of NK-looking cells in the endometrium during a pregnancy.

Conclusions

  • The research thus uncovered the existence of NK-like cell indicator gene expression in horse cells, for the first time ever. This suggested that there is a high prevalence of NK-like cells in the horse endometrium during the early phase of pregnancy.

Cite This Article

APA
Noronha LE, Huggler KE, de Mestre AM, Miller DC, Antczak DF. (2012). Molecular evidence for natural killer-like cells in equine endometrial cups. Placenta, 33(5), 379-386. https://doi.org/10.1016/j.placenta.2012.01.018

Publication

Placenta
ISSN: 1532-3102
NlmUniqueID: 8006349
Country: Netherlands
Language: English
Volume: 33
Issue: 5
Pages: 379-386

Researcher Affiliations

Noronha, L E
  • Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Hungerford Hill Road, Ithaca, NY 14853, USA.
Huggler, K E
    de Mestre, A M
      Miller, D C
        Antczak, D F

          MeSH Terms

          • Amino Acid Sequence
          • Animals
          • Cattle
          • Endometrium / immunology
          • Female
          • Genomics
          • Horses / genetics
          • Horses / immunology
          • Horses / metabolism
          • Humans
          • Killer Cells, Natural / metabolism
          • Lymphocytes / metabolism
          • Mice
          • Molecular Sequence Data
          • Pregnancy
          • Pregnancy, Animal / immunology
          • Pregnancy, Animal / metabolism
          • Receptors, Natural Killer Cell / chemistry
          • Receptors, Natural Killer Cell / genetics
          • Receptors, Natural Killer Cell / metabolism
          • Sequence Homology, Amino Acid
          • Swine

          Grant Funding

          • F32 HD055794 / NICHD NIH HHS
          • R01 HD049545 / NICHD NIH HHS
          • R01 HD049545-05 / NICHD NIH HHS
          • F32 HD 055794 / NICHD NIH HHS

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          Citations

          This article has been cited 11 times.
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