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lijunqing525

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[求助] 英譯漢麻煩幫翻譯下

Due to the complex  interplay  between  platelets  and monocytes,  both  alone and together, during  immune  pathogenesis  of HIV infection, it is essential  to devise  a  better methodology  to detect PMCs. With  this goal  in mind, a  method  to quantitate PMCs in whole blood was standardized  and
was subsequently  employed to assess the percentages of circulating
PMCs in blood collected  from  persons infected with  HIV.
Although  several  previous  reports have  enumerated PMCs using
flow  cytometry in regard  to various  disease  conditions,  the method-ologies  used  thus  far  have  failed to consider  spontaneous  platelet
activation and the resulting  consequence  on  de  novo  PMC  forma-tion (Gkaliagkousi  et al.,  2009; Joseph  et al.,  2001; Ogura et al.,  2001;
Passacquale et al.,  2011; Sevush  et al.,  1998). Thus,  in the efforts  to
formulate an  optimized method  for detection  of PMCs, fixing the
blood samples immediately  following collection was given prime
importance, as  it helped  to conserve  PMCs and avoid experimen-tal  artifacts.  Demonstrated herein,  is a  fix–wash–lyse–wash–stain
method  to fluorescently  label  blood samples,  which when  used
along with  a  progressive  gating strategy and FMO  (fluorescence
minus one)  controls,  allows  for efficient enumeration  of PMCs. The
additional wash  step  performed  after  sample  fixation was neces-sary to avoid the toxicity caused  by  excessive  exposure  to PFA.
One significant concern in adopting  this method  was that the
fixing of samples prior  to staining might  alter  the antibody  binding
capacity to some extent;  however  this was deemed a  reasonable
trade-off  given the advantages  of this method  as  outlined  above.
Nonetheless,  in vitro  whole blood treatments  with  LPS and colla-gen were  performed,  to serve as  monocyte  and platelet activating reagents,  respectively,  in order to ensure  that it is possible to cap-ture the changes in PMC  percentages induced by  these treatments
using  the method  of detection  described here.  Results  indicated
that platelet–monocyte  interaction  increased significantly upon
platelet activation as  compared to monocyte  activation.  These
results  corroborate an  earlier  report by  Rinder  et al.  who  showed
that platelet–leukocyte complexes  increased upon  platelet acti-vation  and that this interaction  was dependent  on  monocyte
activation only  to a  very  limited extent  (Rinder  et al.,  1991).
Upon  validation of the procedure,  persons with  or  without  HIV
infection  (without  any incidence of cardiovascular disease  at least
one year  prior  to enrollment  in the study)  were  enrolled to assess
whether  HIV infection  had any effect on  platelet–monocyte  inter-actions. All  of the individuals  enrolled in the study were  on  ART
and had low viral  load  with  CD4+ T  cell count above 500 cells/mm
3(data not  shown). The results  show that despite  the viral  suppres-sion induced by  successful ART,  the number of circulating  PMCs
was significantly higher  in the inflammatory monocyte  subset  (i.e.
CD16+ monocytes)  of these individuals  as  compared to healthy  indi-viduals  without  HIV infection, and correlated positively with  the
extent  of platelet activation.  The PMC  percentages in the classi-cal  CD16− monocyte  subset  were  also higher  in samples obtained
from  persons infected with  HIV,  although  the effects  were  not  sta-tistically  different  from  controls.  Consistently,  our data  (not shown)
and previous  reports (Pulliam et al.,  1997; Thieblemont  et al.,  1995)
demonstrate  a  significant increase in CD16+ monocyte  percentages
in samples obtained  from  persons infected with  HIV.
This study defines  a  flow  cytometric  method  to quantify  PMCs
in clinical  specimens.  The findings suggest that persons infected
with  HIV have  increased levels of platelet–monocyte  complexes,
particularly  within  the inflammatory monocyte  population  despite
successful ART.  Furthermore,  this data  suggests  that either platelets
have  an  increased preference  to associate  with  CD16+ mono-cytes, or  that alternatively,  the platelet–monocyte  cross-talk causes maturation  of classical monocytes  to the CD16+ phenotype;  thus warranting  further investigation  into whether  this interaction enhances  the monocyte  capacity to cross  the blood brain  barrier, as  well  as  the role of these monocytes  in HIV associated neuro-cognitive  impairment.
Acknowledgements

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寵辱不驚，看庭前花開花落；去留無意，看天外云卷云舒。

1樓 2012-04-10 10:00:52

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xzx8066

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愛與雨下: 金幣+10 2012-04-10 16:43:22
lijunqing525: 金幣+100, 翻譯EPI+1, ★有幫助 2012-04-11 22:37:21

在人類免疫缺陷病毒（HIV）感染的免疫發(fā)病機(jī)制中，血小板和單核細(xì)胞相互作用，無論是單獨(dú)作用還是共同作用，都非常復(fù)雜，因此很有必要建立一個(gè)能更好地檢測(cè)外周血單核細(xì)胞（PMC）的方案。懷著這個(gè)目標(biāo)，我們將一個(gè)定量檢測(cè)全血中PMCs的方法進(jìn)行標(biāo)準(zhǔn)化，隨后應(yīng)用它來評(píng)定HIV感染者血液標(biāo)本中循環(huán)PMCs百分比。
盡管以前有幾個(gè)關(guān)于應(yīng)用流式細(xì)胞儀計(jì)數(shù)不同疾病狀態(tài)PMCs的研究報(bào)道(Gkaliagkousi et al., 2009; Joseph et al., 2001; Ogura et al., 2001; Passacquale et al., 2011; Sevush et al., 1998)，然而他們應(yīng)用的方法不能解決自發(fā)性血小板激活導(dǎo)致的初始PMC形成這個(gè)問題。要構(gòu)想出最佳的檢測(cè)PMCs的方法，血液標(biāo)本采集后迅速固定極其重要，因?yàn)檫@樣有助于保護(hù)PMCs，而且可避免實(shí)驗(yàn)操作導(dǎo)致的人為破壞。
這里介紹的是一種“固定-沖洗-細(xì)胞溶解-沖洗-染色”方法檢測(cè)熒光標(biāo)記血液標(biāo)本。當(dāng)與漸進(jìn)門控技術(shù)和熒光減一控制技術(shù)一起應(yīng)用時(shí)，它可以高效地計(jì)數(shù)PMCs。樣本固定后沖洗這一步驟對(duì)于避免暴露多聚甲醛（PFA）過多導(dǎo)致的毒性是很有必要的。采用這種方法時(shí)一個(gè)非常重要的顧慮就是染色之前固定樣本在某種程度上可能會(huì)改變抗體的結(jié)合能力，然而相對(duì)于這種方法的優(yōu)勢(shì)來說，這可以認(rèn)為是一個(gè)合理的折衷。雖然如此，體外應(yīng)用脂多糖（LPS）和膠原蛋白（作為單核細(xì)胞和血小板的激活劑）分別處理全血標(biāo)本，應(yīng)用這里描述的方法確保能捕捉到這些激活劑誘導(dǎo)的PMC百分比的變化。
結(jié)果顯示，與激活單核細(xì)胞相比，激活血小板可引起血小板-單核細(xì)胞相互作用明顯增加。Rinder等曾研究發(fā)現(xiàn)激活血小板明顯增加血小板-白細(xì)胞復(fù)合體，這種相互作用很少依賴單核細(xì)胞激活(Rinder et al., 1991)。本研究結(jié)果與這是一致的。
為了評(píng)估這種方法的有效性，本研究納入HIV感染和非HIV感染個(gè)體（所有個(gè)體納入研究前，至少1年內(nèi)沒有任何心血管疾病史）評(píng)價(jià)HIV感染是否對(duì)血小板-單核細(xì)胞相互作用有影響。本研究納入的所有個(gè)體均接受抗逆轉(zhuǎn)錄病毒治療（ART），處于低病毒載量且CD4+ T細(xì)胞含量在500個(gè)細(xì)胞/mm3時(shí)期（數(shù)據(jù)未顯示）。
結(jié)果顯示，盡管有效的ART會(huì)導(dǎo)致病毒抑制，與無HIV感染的健康個(gè)體相比，這些個(gè)體循環(huán)PMCs中炎性單核細(xì)胞亞群（如CD16+單核細(xì)胞）數(shù)量明顯增加，并且與血小板激活程度呈正相關(guān)。HIV感染者樣本中經(jīng)典單核細(xì)胞（CD16-）亞群百分比有所增高，然而與對(duì)照組相比無統(tǒng)計(jì)學(xué)差異。一致地，我們的結(jié)果（未顯示）和以前研究（Pulliam et al., 1997; Thieblemont et al., 1995）結(jié)果表明HIV感染者樣本中CD16+單核細(xì)胞百分比明顯增加。
這個(gè)研究精確地描述了一個(gè)量化計(jì)數(shù)臨床樣本中PMCs的流式方法。
即使是在有效的ART情況下，這些結(jié)果顯示HIV感染者血小板-單核細(xì)胞復(fù)合體水平明顯增加，尤其是炎性單核細(xì)胞亞群。此外，這些結(jié)果還顯示血小板優(yōu)先與CD16+單核細(xì)胞增加相關(guān)，或者CD16+單核細(xì)胞優(yōu)先與血小板增加相關(guān)；而且血小板-單核細(xì)胞交互作用促使經(jīng)典單核細(xì)胞轉(zhuǎn)變?yōu)槌墒靻魏思?xì)胞（CD16+）。進(jìn)而，這可確保我們進(jìn)一步研究這種相互作用是否會(huì)增加單核細(xì)胞穿透血腦屏障的能力，以及這些單核細(xì)胞在HIV感染相關(guān)神經(jīng)認(rèn)知損害中作用。致謝！

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2樓2012-04-10 16:15:59

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