综合调查显示，接种COVID-19疫苗后发生了与病毒感染一致的病理生理变化

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综合调查显示，接种COVID-19疫苗后发生了与病毒感染一致的病理生理变化
Comprehensive investigations revealed consistent pathophysiological alterations after vaccination with COVID-19 vaccines
Cell Discovery volume 7, Article number: 99 (2021)
Published: 26 October 2021

　　上海干细胞研究与临床转化研究所、同济大学附属上海东方医院等机构组成的团队对健康成年人接种新型冠状病毒（COVID-19）疫苗进行了一项全面研究。研究结果2021年10月26日发表于 nature（《自然》）杂志。

　　本研究共招募了11名男女健康成年志愿者，年龄24-47岁，BMI为21.5-30.0kg/m2。
　　使用的疫苗为北京生物制品研究所有限公司的灭活SARS-CoV-2疫苗（Vero Cell），肌肉注射到三角肌中。
　　志愿者分为两组；5名参与者（A组）在第1天和第14天接种了全剂量（4μg）灭活SARS-CoV-2疫苗（Vero Cell），6名参与者（B组）在第1天和第28天接种了全剂量疫苗。B组中的一名志愿者在接种疫苗前被检测出抗SARS-CoV-2 IgM和IgG呈阳性，但没有关于COVID-19核酸诊断阳性的记录，这表明潜在的先前感染。
　　该研究表明，健康志愿者接种灭活SARS-CoV-2疫苗后，除了产生中和抗体外，糖化血红蛋白、血清钠和钾水平、凝血曲线和肾功能发生持续变化。在COVID-19患者中也报告了类似的变化，这表明疫苗接种模仿了感染。第一次接种前和接种后28天外周血单核细胞(PBMC)的单细胞mRNA测序（scRNA-seq）也揭示了许多不同免疫细胞类型的基因表达的一致变化。减少CD8+T细胞和经典单核细胞含量的增加是典型的。此外，scRNA-seq显示NF-κB信号增加和I型干扰素反应减少，这已通过生物测定得到证实，并且据报道在SARS-CoV-2感染后症状加重。总之，该研究建议在为患有糖尿病、电解质失衡、肾功能不全和凝血障碍的人接种疫苗时要格外小心。
  Here, we report, besides generation of neutralizing antibodies, consistent alterations in hemoglobin A1c, serum sodium and potassium levels, coagulation profiles, and renal functions in healthy volunteers after vaccination with an inactivated SARS-CoV-2 vaccine. Similar changes had also been reported in COVID-19 patients, suggesting that vaccination mimicked an infection. Single-cell mRNA sequencing (scRNA-seq) of peripheral blood mononuclear cells (PBMCs) before and 28 days after the first inoculation also revealed consistent alterations in gene expression of many different immune cell types. Reduction of CD8+ T cells and increase in classic monocyte contents were exemplary. Moreover, scRNA-seq revealed increased NF-κB signaling and reduced type I interferon responses, which were confirmed by biological assays and also had been reported to occur after SARS-CoV-2 infection with aggravating symptoms. Altogether, our study recommends additional caution when vaccinating people with pre-existing clinical conditions, including diabetes, electrolyte imbalances, renal dysfunction, and coagulation disorders.

　　【研究结果】

　　一、总体而言，接种后的不良反应是轻微的（1级或2级）和短暂的。
  Overall, adverse reactions were mild (grades 1 or 2) and transient.

　　二、接种灭活SARS-CoV-2疫苗后的抗SARS-CoV-2抗体和中和抗体
　　A组的检测结果表明，在第2次接种之前，0%的参与者产生了抗SARS-CoV-2 IgG，但到第28天，即第2次接种后2周，100%的参与者检测为阳性。总体而言，IgM比IgG出现得更早，这是意料之中的。A组的IgG和IgM阳性在第42天下降，到第90天仍保持在相对较低的水平。
　　对于B组，直到第2次接种后才出现IgG。然而到第42天，IgG阳性达到100%并持续到第56天，这表明B组的疫苗接种方案更有效。到第90天，IgG阳性率也降低到50%，表明抗体生产无法长时间维持。
　　进一步对SARS-CoV-2中和抗体进行测试的结果还表明，与相隔14天的两次接种（A组）相比，相隔28天的两次接种（B组）产生更高的保护性抗体滴度。另一方面，如之前报道的，抗SARS-CoV-2中和抗体滴度总体上低于COVID-19恢复期个体。到90天，所有志愿者的中和抗体滴度均显著下降。有趣的是，与其他参与者相比，在疫苗接种前抗体呈阳性的个体并不容易产生中和抗体，这表明先前潜在的感染可能没有发生，或者从中和抗体产生的角度来看可能不会产生长期的保护。
  Testing results from cohort A demonstrated that prior to the 2nd inoculation 0% of the participants developed anti-SARS-CoV-2 IgG, but by day 28, which was 2 weeks post the 2nd inoculation, 100% of the participants were tested positive. Overall, IgM showed up earlier than IgG, which was expected. IgG and IgM positivity decreased by day 42 and remained at relatively low levels by day 90 in cohort A. For cohort B, no one developed IgG until after 2nd inoculation. Yet by day 42, IgG positivity reached 100%  and sustained until day 56, suggesting that the vaccination protocol for cohort B was more efficacious. By day 90, IgG positivity also reduced to 50%, indicating antibody production did not sustain for a long time. We further carried out tests for SARS-CoV-2 neutralizing antibodies18, and results also indicated that two inoculations 28 days apart (cohort B) resulted in higher protective antibody titers as compared to two inoculations with 14 days apart (cohort A). On the other hand, it appeared that anti-SARS-CoV-2 neutralizing antibody titers were overall lower than those in COVID-19 convalescent individuals as reported before3. By 90 days, neutralizing antibody titers dramatically decreased in all volunteers. Interestingly, the individual who was antibody positive prior to vaccination was not more prone to generating neutralizing antibodies as compared to the rest of the participants, suggesting that prior potential infection might not have occurred or may not generate long-lasting protection in the perspective of neutralizing antibody production.

　　三、疫苗接种后临床实验室测量值的变化
　　在接种疫苗后第7天，白细胞计数显著增加，但仅略有增加。在其后的时间点未检测到差异。
　　在健康志愿者中观察到糖化血红蛋白（HbA1c）水平相当一致的增加，无论他们属于A组还是B组。到第1次接种后第28天，11人中有3人达到糖尿病前期范围。到第42天和第90天，中等HbA1c水平似乎恢复了，但仍显著高于接种疫苗前的水平。先前的研究表明，血糖水平不受控制的糖尿病患者更容易患上严重的COVID-19。高血糖水平/糖酵解已被证明可通过线粒体活性氧的产生和HIF1A的激活来促进人类单核细胞中SARS-CoV-2的复制，因此呈现出不利的特征。
　　第一次接种后第28、42和90天血清钾水平显著下降，其中一个样本在第42天低于正常下限。同样，接种疫苗后血清钠水平也降低，表明疫苗对电解质平衡有影响。同样，电解质失衡也与COVID-19有关。
　　凝血病是另一种由COVID-19引起的临床病症。研究发现接种疫苗后凝血曲线发生了显著变化，在第一次接种后的短期（7天）内，凝血曲线倾向于较短的凝血酶原时间（PT），而长期（28天和42天）效果则倾向于活化部分促凝血酶原激酶时间（APTT）和PT延长。到第90天，这些特征恢复到接种疫苗前的状态。
　　研究还发现第一次接种后第7天、第28天血液胆固醇水平升高，第7天也检测到总胆汁酸水平升高。
　　肾功能不全是与COVID-19相关的另一种临床病症，在第一次接种后28、42和90天，血清肌酐水平显著高于接种前，导致估算肾小球滤过率（eGFR）降低。
　　据报道，这些临床特征中的大多数与COVID-19患者出现严重症状有关。
  White blood cell count was significantly, yet only slightly, increased after vaccination on day 7. No differences were detectable at the following time points. To our surprise, quite consistent increases in HbA1c levels were observed in healthy volunteers, regardless of whether they belonged to cohort A or B. By day 28 post the 1st inoculation, three out of 11 individuals reached the prediabetic range. By days 42 and 90, medium HbA1c levels appeared to revert back, yet were still significantly higher than those before vaccination. Previous work has demonstrated that diabetic patients with uncontrolled blood glucose levels are more prone to develop severe forms of COVID-19. High blood glucose levels/glycolysis had been shown to promote SARS-CoV-2 replication in human monocytes via the production of mitochondrial reactive oxygen species and activation of HIF1A, therefore presenting a disadvantageous feature.
  Serum potassium levels decreased significantly by days 28, 42, and 90 post the 1st inoculation, with one sample below the lower normal limit at day 42. Similarly, serum sodium levels also decreased following vaccination, indicative of vaccine influences on electrolyte balance. Again, electrolyte imbalance has also been linked to COVID-19. Coagulopathy is another COVID-19-induced clinical condition. We found that coagulation profiles changed significantly after vaccination, in the short-term (7 days) after the 1st inoculation, coagulation profiles were leaning toward shorter Prothrombin Time (PT), whereas the long-term (28 and 42 days) effect was toward activated partial thromboplastin time (APTT) and PT prolongation. By day 90, the profiles returned back to those before vaccination. Moreover, we found elevated blood cholesterol levels at days 7, 28 after the 1st inoculation, and elevated total bile acid levels were also detected at day 7. Renal dysfunction is another clinical condition linked to COVID-19, and by 28, 42, and 90 days after the first inoculation, serum creatinine levels were significantly higher than those before vaccination, resulting in reduced eGFR. Most of these clinical features have been reported to be associated with the development of severe symptoms in COVID-19 patients.

　　四、单细胞mRNA测序（scRNA-seq）揭示了疫苗接种后几乎所有免疫细胞的基因表达发生了巨大变化
　　关于疫苗接种前后细胞类型组成的差异，观察到接种疫苗后CD4+调节性T细胞(CD4.Treg)、CD8+T细胞(CD8.T)和增殖性CD8+细胞(CD8.Tprolif)的含量降低。γδ-T细胞（gd.T.Vd2）含量的减少也很显著。相比之下，疫苗接种增加了CD14+经典单核细胞(Mono.C)含量，与临床实验室测量结果一致。包括所有CD4+T细胞、所有CD8+T细胞、B细胞和NK细胞在内的总淋巴细胞含量在接种疫苗前后没有显著变化，这也通过临床实验室测量得到证实。从196名COVID-19感染患者和对照中收集已发布的数据集分析表明，与对照组相比，在COVID-19患者中，除了增殖的CD8+T细胞，疫苗引起的所有五种不同免疫细胞亚型的细胞含量变化也发生了相同的变化。
　　关于疫苗接种引起的详细基因表达变化，研究发现显著上调的基因参与“通过NF-κB的TNFα信号传导”、“炎症反应”和“细胞因子-细胞因子受体相互作用”、“IL6-JAK STAT3信号传导”、“凝血”、“缺氧”，而在关于COVID-19的报道中，细胞周期相关途径被下调。这些结果支持了疫苗接种模拟感染的观点。
  To reveal differences in cell-type compositions before and after vaccination, we calculated relative percentages of all cell types in PBMCs of each individual on the basis of scRNA-seq data. We observed decreases in contents of CD4+ regulatory T cells (CD4.Treg), CD8+ T cells (CD8.T), and proliferating CD8+ cells (CD8.Tprolif) after vaccination. Decreases in γδ-T cell (gd.T.Vd2) contents were also significant. In contrast, vaccination increased CD14+ classical monocyte (Mono.C) contents, consistent with clinical laboratory measurements. The overall lymphocyte contents, which included all CD4+ T cells, all CD8+ T cells, B cells, and NK cells, did not change significantly before and after vaccination, which was also confirmed by clinical laboratory measurements. We collected a published dataset from 196 COVID-19-infected patients and controls7, and analyzed our data together with that dataset. The result indicated that vaccination-induced changes in cell contents of all five different immune cell subtypes also changed in the same directions in COVID-19 patients as compared to controls, except for proliferating CD8+ T cells.
  To study detailed gene expression changes induced by vaccination, we merged individual samples into pseudo-bulk samples and used paired sample test to identify differentially expressed genes (DEGs). Significantly upregulated genes were involved in “TNFα signaling via NF-κB”, “inflammatory responses”, and “cytokine-cytokine receptor interaction”, “IL6-JAK STAT3 signaling”, “coagulation”, “hypoxia”, which had been reported for COVID-19, while cell cycle-related pathways were downregulated. These results supported the notion that vaccination mimicked an infection.

　　五、特色免疫细胞亚型特异性基因表达变化反映了临床实验室的变化
　　在所有主要细胞类型中鉴定了差异表达基因（DEG）并进行了基因功能分析。
　　与临床测量结果相呼应的是，与“胆固醇稳态”、“凝血”和“炎症反应”相关的基因（CXCL8、CD14、IL6和TNFRSF1B）、“TNFα通过NF-κB信号传导”（NFKB1、NFKB2、NFKBIE、TNFAIP3,和TNFSF9)和“缺氧”(HIF1A)被上调。此外，“TGFβ信号”、“IL2-STAT5信号”（IFNGR1、MAPKAPK2和CASP3）和“IL6-JAK-STAT3信号”相关基因也上调。
　　有趣的是，“炎症反应”基因在单核细胞中高度表达，并且在接种疫苗后进一步增加，表明单核细胞是接种疫苗后参与炎症反应的主要细胞类型之一。相比之下，与“糖酵解”、“胆汁酸代谢”和“I型干扰素(IFN-α/β)反应”相关的基因被下调，与我们的临床数据和COVID-19的病理生理学一致。
  We identified diferentially expressed genes (DEGs) among all major cell types and conducted gene functional analysis. Echoing the clinical measurement results, genes related to “cholesterol homeostasis”, “coagulation”, and “inflammatory response” (CXCL8, CD14, IL6, and TNFRSF1B), “TNFα signaling via NF-κB” (NFKB1, NFKB2, NFKBIE, TNFAIP3, and TNFSF9) and “hypoxia” (HIF1A) were upregulated. In addition, “TGFβ signaling”, “IL2-STAT5 signaling” (IFNGR1, MAPKAPK2, and CASP3), and “IL6-JAK-STAT3 signaling”-related genes were also upregulated. To visualize which cell types were enriched for those signatures, we performed gene module scoring and displayed the scores on UMAP coordinates as well as grouped box plots. Interestingly, “inflammatory response” genes were highly expressed in monocytes and after vaccination further increased, suggesting monocytes were one of the major cell types participating in inflammatory responses after vaccination. In contrast, genes related to “glycolysis”, “bile acid metabolism”, and “type I interferon (IFN-α/β) response” were downregulated, consistent with our clinical data and the pathophysiology of COVID-19.

　　六、多种免疫细胞亚型的最常见变化显示NF-κB信号传导增加和IFN-α/β反应减少
　　鉴定出接种后排名最高（最活跃）的八个上调的调节子和八个下调的调节子。我们选择了3+3个典型的调节子来构建一个调节网络，该网络显示了两个不同的组，一个由接种后下调的IRF2、STAT1和STAT2组成，另一个包含接种后上调的RELB、NFKB2和HIF1A。上调网络的GO项主要与淋巴细胞分化、活化和“生发中心形成”有关，这表明T细胞和B细胞在接种疫苗后被激活。此外，接种疫苗后NF-κB信号传导也升高。下调的网络富含许多干扰素相关途径和细胞因子分泌。这表明疫苗接种可能通过降低调节子STAT1、STAT2和IRF2的活性来抑制外周免疫系统中的干扰素反应，这些调节子被认为是驱动I型和III型干扰素信号传导的主要转录因子。
　　为了证实scRNA-seq揭示的疫苗接种诱导的干扰素反应抑制，在接种IFN-α/β疫苗之前和之后28天刺激了接种疫苗个体的外周血单核细胞（PBMC）。在培养16小时和刺激12小时后，使用RT-qPCR测量主调节因子IRF2、IRF7和STAT2的相对表达。接种后STAT2和IRF7显著下调，而IRF2呈现下调趋势。调节子分析表明，接种疫苗后外周免疫系统的状态降低了I型干扰素反应，表明在第一次接种后至少28天，一般抗病毒能力减弱。
  Top-ranked (most active) eight regulons upregulated and eight regulons downregulated after vaccination were identified. We selected 3+3 typical regulons to construct a regulatory network as presented in Fig. 5c. The network showed two distinct groups, one is consisted of IRF2, STAT1 and STAT2, which were downregulated after vaccination, and the other, contained RELB, NFKB2, and HIF1A, which were upregulated after inoculation. The GO terms of the upregulated network are predominantly related to lymphocyte differentiation, activation, and “Germinal Center Formation”, which suggested that T cells and B cells were activated after vaccination. In addition, NF-κB signaling was also elevated after vaccination. The downregulated network was enriched for many interferons-related pathways and Cytokine Secretion. This suggested that vaccination might inhibit interferon responses in the peripheral immune system, by reducing the activities of regulons STAT1, STAT2, and IRF2, which were thought to be master transcription factors driving type I and III interferon signaling.
  To confirm vaccination-induced inhibition of interferon responses revealed by scRNA-seq, we stimulated PBMCs from vaccinated individuals before and 28 days after vaccination with IFN-α/β. After 16 h of culturing and 12 h of stimulation, we used RT-qPCR to measure the relative expression of master regulators IRF2, IRF7, and STAT2. STAT2 and IRF7 were significantly downregulated after vaccination, yet IRF2 showed a trend of downregulation. The regulon analyses indicated that the states of the peripheral immune system after vaccination had reduced type I interferon responses, indicative of attenuated general antiviral abilities at least 28 days after the first inoculation.

　　七、单核细胞中疫苗接种诱导的炎症反应
　　最近的报告描述了对呼吸道病毒感染的保守宿主免疫反应特征，即元病毒特征（MVS），它在SARS-CoV-2感染中也是保守的。较高的MVS评分与感染相关。研究发现，接种疫苗后MVS评分显著更高，表明疫苗接种模拟了感染。有趣的是，阳性MVS基因组主要在单核细胞中表达，而阴性组在淋巴细胞中，表明接种疫苗后会发生不同的细胞类型特异性免疫反应。
　　研究发现，与MVS评分和MVS阳性组最高度相关的途径是“炎症反应信号”，在接种疫苗后，它与CD14、FPR1、C5AR1、NAMPT、NLRP3、CDKN1A和IFNGR2在单核细胞中显著上调。然而，MVS阴性组与“细胞毒性特征”密切相关，以NKG7、CCL4、CST7、PRF1、GZMA、GZMB、IFNG和CCL3表达为代表，在许多T细胞亚型中显著降低，但在接种疫苗后NK细胞没有显著降低。
Recent reports have described conserved host immune response signatures to respiratory viral infections, namely the Meta-Virus Signature (MVS), which is also conserved in SARS-CoV-2 infection. Higher MVS scores are associated with infection. In all, 380 (158 positively- and 222 negatively contributed to MVS scores) out of 396 (161 positively- and 235 negatively contributed) genes selected for MVS measurement were detected in our dataset. To investigate host immune responses after vaccination with inactivated SARS-CoV-2, we separated the positive and negative gene sets and calculated MVS scores. The MVS scores were substantially higher after vaccination, suggesting that vaccination mimicked an infection. Interestingly, the positive MVS gene set was predominantly expressed in monocytes, while the negative set in lymphocytes, indicating different cell-type-specific immune responses would take place after vaccination.
  The most highly correlated pathway with MVS score and MVS-positive set was “Inflammatory response signaling”, which was strikingly upregulated in monocyte after vaccination, together with CD14, FPR1, C5AR1, NAMPT, NLRP3, CDKN1A, and IFNGR2. Whereas, MVS-negative set correlated well with “Cytotoxicity signature”, represented by NKG7, CCL4, CST7, PRF1, GZMA, GZMB, IFNG, and CCL3 expression, significantly decreased in many T-cell subtypes but not NK cells after vaccination.

　　【讨论】

　　这是对病理生理变化的全面调查，包括接种COVID-19后人们的详细免疫学变化。结果表明，疫苗接种除了刺激中和抗体的产生外，还影响了各种健康指标，包括与糖尿病、肾功能障碍、胆固醇代谢、凝血问题、电解质失衡有关的指标，就像志愿者经历了感染一样。疫苗接种前后志愿者PBMC的scRNA-seq显示免疫细胞基因表达发生了巨大变化，这不仅与一些临床实验室测量结果相呼应，而且表明NF-κB相关炎症反应增加，结果证明这主要发生在在经典的单核细胞中。疫苗接种也增加了经典的单核细胞含量。此外，对MVS评分有积极贡献的基因组（也已知与严重症状发展相关）在单核细胞中高度表达。据称对COVID-19有益的I型干扰素(IFN-α/β)反应在接种疫苗后被下调。此外，阴性MVS基因在淋巴细胞（T、B和NK细胞）中高表达，但在接种疫苗后表达降低。总之，这些数据表明，在接种疫苗后，至少到第28天，除了产生中和抗体外，人们的免疫系统，包括淋巴细胞和单核细胞的免疫系统，可能处于更脆弱的状态。
This is a comprehensive investigation of the pathophysiological changes, including detailed immunological alterations in people after COVID-19 vaccination. Results indicated that vaccination, in addition to stimulating the generation of neutralizing antibodies, also influenced various health indicators including those related to diabetes, renal dysfunction, cholesterol metabolism, coagulation problems, electrolyte imbalance, in a way as if the volunteers experienced an infection. scRNA-seq of PBMCs from volunteers before and after vaccination revealed dramatic changes in immune cell gene expression, not only echoing some of the clinical laboratory measures but also suggestive of increased NF-κB-related inflammatory responses, which turned out to be mainly taking place in classical monocytes. Vaccination also increased classical monocyte contents. Moreover, the gene set positively contributing to MVS scores, also known to be associated with severe symptom development, was highly expressed in monocytes. Type I interferon (IFN-α/β) responses, supposedly beneficial against COVID-19, were downregulated after vaccination. In addition, the negative MVS genes were highly expressed in lymphocytes (T, B, and NK cells), yet showed reduced expression after vaccination. Together, these data suggested that after vaccination, at least by day 28, other than generation of neutralizing antibodies, people’s immune systems, including those of lymphocytes and monocytes, were perhaps in a more vulnerable state.

　　有趣的是，我们的初步数据表明，如果我们将SARS-CoV-2的RBD与PBMC（来自接种疫苗前后的志愿者）预孵育，然后用IFN-α/β处理细胞，实际上I型干扰素反应在接种疫苗后的PBMC，这表明可能接种疫苗虽然降低了一个人的一般抗病毒能力，但增强了特异性针对SARS-CoV-2的适应性免疫功能。另一方面，比较疫苗接种前的PBMC，SARS-CoV-2S-RBD的预处理似乎降低了I型干扰素反应（P＜0.05，IRF2，IRF7，STAT2），表明第一次接触病毒肽实际上会导致PBMC中I型干扰素反应的减少。这些体外数据很好地支持了scRNA-seq结果。
  Interestingly, our preliminary data demonstrated that if we pre-incubated RBD of SARS-CoV-2 with the PBMCs (from volunteers before and after vaccination) and then treated the cells with IFN-α/β, type I interferon responses were actually enhanced in PBMCs after vaccination, suggesting that perhaps vaccination, while reduced a person’s general antiviral ability, enhanced adaptive immune function specifically towards SARS-CoV-2 . On the other hand, comparing PBMCs before vaccination, pre-treatment of SARS-CoV-2 S-RBD appeared to reduce type I interferon responses (P<0.05, IRF2, IRF7, STAT2), suggesting 1st time exposure of the viral peptide would actually cause a reduction in type I interferon responses in PBMC. These in vitro data nicely supported the scRNA-seq results.

　　值得一提的是，A组中一个服用抗生素的人碰巧没有降低与I型干扰素反应相关的基因表达，并且该人在该组中也具有最高的中和抗体滴度。我们进一步计算了中和抗体滴度与炎症反应之间的Pearson相关系数，通过NF-κB和干扰素-α（I型干扰素）反应与TNFα信号传导相关基因的平均基因表达测量。结果为0.32和0.39，P＞0.05，分别表明疫苗的免疫反应变化和适应性免疫保护似乎并不高度相关。抗生素是否会影响疫苗效力仍有待确定。同样有趣的是，虽然A组和B组具有不同的抗SARS-CoV-2抗体产生谱，但他们的PBMC sscRNA-seq结果非常相似，包括他们的B细胞scRNA-seq数据。需要注意的是，接种疫苗后，大多数反应性B细胞，尤其是产生成熟抗COVID-19抗体（IgG）包括记忆B细胞的B细胞，应主要位于淋巴结和脾脏等外周淋巴组织中，而只有少数成熟的B细胞存在于循环中。因此，PBMCs制剂中的B细胞群可能无法反映体液免疫的全谱。
It is worth mentioning that one individual in cohort A who was on antibiotics, happened to not having reduced gene expression linked to type I interferon responses, and this individual also had the highest neutralizing antibody titer within the cohort. We further calculated Pearson’s Correlation Coefficient between neutralizing antibody titers and inflammatory responses measured by averaged gene expression of genes associated with TNFα Signaling via NF-κB and interferon-α (type I interferon) responses. The results were 0.32 and 0.39 with P>0.05, respectively, suggesting immune response changes and adaptive immune protection of the vaccine do not appear to be highly correlated. Whether antibiotics may influence vaccine efficacy remains to be determined. It is also rather interesting that while cohorts A and B had different anti-SARS-CoV-2 antibody production profiles, their PBMCs scRNA-seq results were drastically similar, including their B-cell scRNA-seq data. It should be noted that after vaccination, the majority of responsive B cells, particularly those producing mature anti-COVID-19 antibodies (IgG) including memory B cells, should be primarily located in peripheral lymphatic tissues such as lymph nodes and the spleen, while only a few mature B cells would exist in the circulation. Therefore, the B-cell population in PBMCs preparations may not reflect the whole spectrum of humoral immunity.

　　本研究中提出的分析，特别是PBMC的scRNA-seq尚未针对之前的疫苗评估进行过，无论免疫系统功能相关基因的变化是COVID-19特异性还是可以普遍应用于其他疫苗或其他类型COVID-19疫苗的数量仍有待确定。然而，这些类型的详细分析应该总体上有益于疫苗的开发和应用。我们的研究假设，必须考虑疫苗接种对某些医疗条件或对一般人类健康的潜在长期影响。
  The analyses presented in this study, particularly, scRNA-seq of PBMCs had not been performed for previous vaccine evaluations, whether the changes in immune system function-related genes were COVID-19-specific or could be generally applied to other vaccines or other types of COVID-19 vaccines remained to be determined. However, these types of detailed analyses should be overall beneficial to vaccine development and applications. Our study postulates that it is imperative to consider the potential long-term impact of vaccination to certain medical conditions or to general human health.

本文仅供学习参考，完整准确内容请查阅原始文献：Liu, J., Wang, J., Xu, J. et al. Comprehensive investigations revealed consistent pathophysiological alterations after vaccination with COVID-19 vaccines. Cell Discov 7, 99 (2021). https://doi.org/10.1038/s41421-021-00329-3
原始地址：https://www.nature.com/articles/s41421-021-00329-3