Korean J Obstet Gynecol.  2011 Jul;54(7):361-368. 10.5468/KJOG.2011.54.7.361.

In vitro suppression of natural killer cell cytolytic activities by prednisolone in patients with history of unexplained recurrent spontaneous abortion and elevated peripheral blood NK cell fraction

• Ahn HS
• Cha SH
• Koo HS
• Bae JY
• Yoo JH
• Song IO
• Ahn HK
• Hwang SJ
• Park DW
• Yang KM

• KMID: 2274050
• DOI: http://doi.org/10.5468/KJOG.2011.54.7.361

Abstract

OBJECTIVE
We aimed to evaluate the effect of prednisolone (PDS) on natural killer cell (NK cell) cytolytic activity in vitro.
METHODS
Blood samples from 74 patients with history of unexplained recurrent spontaneous abortion who elevated peripheral blood NK cell fraction were collected prospectively. Peripheral blood monocytes which containing NK cells were isolated and separated to three different tubes which containing target (K562) cells by the 50:1 effector to target (E:T) ratio. PDS or intravenous immunoglobulin (IVIG) was additionally added to 2 tubes for evaluate their suppressive effect. The percentage killing of target cells was recorded numerically by using flow cytometer and the values between groups were statistically analyzed.
RESULTS
The mean target cell killing percentage was 40.5% in co-culture tube which was not added PDS or IVIG. In culture experiments which was added IVIG, the killing percentage is reduced to 37.7% which showed no significant differences compared to that of co-cultured tube which was not added PDS or IVIG. But, in experiments with added PDS, the killing percent was reduced to 19.5% and the difference was statistically significant (P<0.001) compared to that of co-cultured tube which was not added PDS or IVIG. On comparing the reduction in killing percentage of target cells by PDS and IVIG, statistically significant reduction in the PDS coculture was noted (P<0.005).
CONCLUSION
NK cell cytolytic activity is effectively down-regulated by using PDS in vitro. Moreover, the effect of PDS in down-regulation of NK cell cytolytic activity is seems to superior than that of IVIG. But, large scaled in vivo study is needed.

Keyword

Recurrent miscarriage; Natural killer cell cytolytic activity; Prednisolone

MeSH Terms

Abortion, Habitual
Abortion, Spontaneous
Coculture Techniques
Down-Regulation
Female
Homicide
Humans
Immunoglobulins
Immunoglobulins, Intravenous
Killer Cells, Natural
Monocytes
Prednisolone
Pregnancy
Prospective Studies
Immunoglobulins
Immunoglobulins, Intravenous
Prednisolone

Figure

• 

  Fig. 1 Determination of immune cell profiles in peripheral blood. Gating strategy for three color flow cytometry evaluation of peripheral blood immune cell. This step descript of analysis gate around lymphocyte (R1), excluding cell debris (A). Next step shows four regions as NK cell; CD3-/16+/56+. (B) NKT cell; CD3+/16+/56+, T cell; CD3+/56- and B cell (CD3-/56-) each. SSC, side scatter; PI, propidum iodie dye; PKH2, green fluorescent cell linker dye; NKT, natural killer T cell; NK, natural killer.

• 

  Fig. 2 Determination of peripheral blood natural killer cell cytolytic activities. Figures describes analysis gate around K562 (target cell) live & dead cell in control (A, no effector cell), 50:1 effector cell:target cell (E:T) ratio (B), 25:1 E:T ratio (C), and 12.5:1 ET ratio (D) each which excluding cell debris. PKH2, green fluorescent cell linker dye; PI, propidum iodie dye.

• 

  Fig. 3 Mean percentage of target cell killed with different effector to target cell ratios. aP<0.05, analysis of variance.

• 

  Fig. 4 Mean percent to target cells with intravenous immunoglobulin or prednisolone coculture. IVIG, intravenous immunoglobulin; PDS, prednisolone. aP<0.05, analysis of variance (ANNOVA). bP<0.05, ANNOVA.

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