Protection of Lethally Irradiated Mice by Spleen Cells from Neonatally Thymectomized Mice

Long-lived, immunologically vigorous (C3H(f) x A(f))F(1) hybrids were produced after lethal irradiation by administration of spleen cells from C3H(f) or syngeneic donors. Further, neonatally thymectomized C3H(f) or A(f) strain donors reconstituted irradiated C3H(f) or (C3H(f) x A(f))F(1) hosts. In addition, C3H(f) spleen cells from nonthymectomized 10- to 15-day-old donors protected irradiated hybrid mice, but A(f) cells of young mice as well as of older mice produced graft-versus-host reaction and early death in irradiated C3H(f) or (C3H(f) x A(f))F(1) hybrids.Abrogation of secondary disease by treatment of irradiated mice with spleen cells from allogeneic neonatally thymectomized mice is possibly attributable to diminished immunologic competence of the cells grafted, followed by the development of immunological tolerance of the donor cells. Donor cells, receiving thymus influence in the recipient host after transplantation, could explain the long-lived immunologically vigorous radiation chimeras that did not experience graft-versus-host reactions. The findings of this study help to understand the differential susceptibility of A(f) and C3H(f) mice to development of tolerance to one another's antigens observed in prior investigations.It appears that, in these mice, the host thymus influences the maturation of the spleen cells from young mice or from neonatally thymectomized mice. However, this influence was often greater in mice given 767 rads than in those given 1046 rads. This differential influence is possibly attributable to irradiation damage to the thymus produced by the higher dose of irradiation. Spleen cells from neonatally thymectomized mice can be differentiated and expanded by the thymus of the host. The differential susceptibility of T(1), early differentiation stages, of thymus-dependent lymphocytes and T(2), late differentiation stages, of thymus-dependent lymphocytes to tolerance induction and immunostimulation, respectively, are proposed as the bases for these otherwise paradoxical influences.     

Non-Pathogenic Bacteria Modulate Colonic Epithelial Gene Expression in Germ-Free Mice

Background: We established a bacterial reconstitution model to investigate epithelial cell-luminal bacteria interaction. The aim of the study was to identify the known genes directly or indirectly modulated by non-pathologic bacterial flora in the colonic epithelia of germ-free mice. Methods: Germ-free mice were orally given a bacterial suspension prepared from specific pathogen-free counterparts (bacterial reconstitution). Colonic epithelial cells were isolated, then total and poly (A) RNA were extracted. We investigated differential gene expression in colonic epithelial cells among germ-free, bacteria-reconstituted, and specific pathogen-free mice by DNA microarray. Finally, differential expression was confirmed by Northern blot or quantitative RT-PCR. Results: Thirty genes were initially selected as differentially expressed genes in DNA microarray analysis. We confirmed that genes associated with growth (Reg III &#35, Reg III &#37 , guanylate nucleotide binding protein 2), apoptosis (Bcl-associated death promotor), cytoskeleton (tubulin &#33 4, erythrocyte protein band 7.2), and immune response (lymphocyte antigen complex 6) were induced by bacterial reconstitution. In contrast, genes possibly participating in extracellular oxidant defence (selenoprotein P, metallothionein 1) and cellular metabolism (cytochrome P450, HMGCoA synthase 2, alcohol dehydrogenase 1 complex, aldehyde dehydrogenase family 1, carbonic anhydrase 1, glycoprotein galactosyltransferase &#33 1,3) were down-regulated by bacterial challenge. Conclusion: Non-pathogenic bacteria modulated colonic gene expression in germ-free mice, suggesting that non-pathogenic bacteria possibly initiate epithelial change in genetically normal and/or abnormal hosts. The present study provides a basis for the functional study of each molecule in symbiosis with luminal bacteria in healthy and diseased colon.     

Gene-Targeted Mice in Leukocyte Adhesion Research

Physiologic and inflammatory leukocyte trafficking are controlled by adhesion molecules on leukocytes and endothelial cells. These adhesion molecules can be grouped into four major families: integrins, immunoglobulinlike molecules, selectins, and glycoproteins serving as selectin ligands. The recent advent of gene-targeting technology in embryonal stem cells has prompted the production of mutant mice that lack individual adhesion molecules or combinations thereof. Such gene-targeted mice permit studies into the roles of adhesion molecules in acute and chronic inflammation and of the regulation and interplay between different sets of adhesion receptors in vivo. Microcirculatory studies have been indispensable to our understanding of the importance of certain adhesion molecules for different steps of leukocyte recruitment in vivo. Targeting new genes and investigating mice with combined adhesion-molecule deficiencies will help to further clarify the molecular mechanisms of leukocyte trafficking in health and disease. This article reviews the insight gained through using adhesion molecule-deficient mice, emphasizing the role of microcirculatory studies in this research.     

Lower Total Leukocyte and Neutrophil Counts in Healthy Young Africans from Uganda

The total leukocyte count (TLC) is an important component of the complete blood count and influences many clinical decisions. The effect of race or ethnicity on TLC is not well known. The African population has been reported to have lower than normal TLC and neutrophil counts. In this study, thirty eight African students referred for medical check up to a tertiary care hospital were included. Complete blood count was done on a three part automated hematology analyzer. Blood smear examination and manual differential count was also done. The control group included 38 age and sex matched healthy individuals. Student t test was used to compare the differences between the groups. The mean TLC in African students (4.95 ± 1.09 × 10⁹/l) was significantly lower (p < 0.0001) than that seen in the control group (7.42 ± 1.7 × 10⁹/l). The mean neutrophil percentage was also lower (49 ± 7.5%) in African students compared to the control group (63.6 ± 9.8%) [p < 0.0001] while lymphocyte percentage was higher (45.2 ± 7.5%) in the African students as compared to the control group (31.0 ± 9.3%) [p < 0.0001]. Absolute neutrophil count was also lower (2.45 ± 0.76 × 10⁹/l) in African students compared to the control group (4.76 ± 1.47 × 10⁹/l) while absolute lymphocyte count was comparable (2.21 ± 0.5 × 10⁹/l vs. 2.26 ± 0.72 × 10⁹/l) [p = 0.7212]. This study has shown lower leukocyte and neutrophil counts in apparently healthy African individuals. Knowledge of this variation from normal white cell and neutrophil counts is important in avoiding unnecessary investigations and influencing therapeutic decisions in these individuals.     

The Effect of Orally Administered Bovine Spleen Preparations on Platelet and Leukocyte Counts of Rats

Several orally administered beef spleen preparations were evaluated fortheir effect on platelet and leukocyte counts of rats. In some studies, cowspleen exerted a definite though variable count lowering activity. Furtherstudies yielded less dramatic or negative results. Pooled beef spleen, wholeor crudely fractionated, had no demonstrable effect on the blood counts.

Other variables, including temperature, feeding methods, seasonal factors,and animal housing did not appear significant.

No consistent, predictable effect of beef spleen feedings could be demonstrated under the various conditions tested.

Submitted on January 9, 1964 Accepted on March 21, 1965     

Fluctuations in the Peripheral Blood Leukocyte and Platelet Counts in Leukocytapheresis in Healthy Volunteers

Abstract: Leukocytapheresis (LCP) for the treatment of patients with diseases that involve an abnormal autoimmune reaction aims to improve the condition of the patient's pathology and to correct imbalances in immunological regulation mechanisms by removing the responsible leukocytes from the peripheral blood. To clarify the mechanism of therapeutic effect, LCP was conducted in healthy volunteers to investigate changes in peripheral blood leukocyte and platelet counts over time during the treatment. The subjects were 10 healthy male volunteers. LCP was performed once in each volunteer for 3,000 ml of blood volume. The peripheral blood counts decreased significantly, reaching a minimum of 20.0% of the baseline number of leukocytes, 10.1% of the baseline number of neutrophils, and 40.3% of the baseline number of lymphocytes. The number of removed leukocytes was about 6.6 × 10⁹ cells, including about 3.5 × 10⁹ neutrophils, as well as about 5.0 × 10¹¹ platelets. After the completion of LCP, the peripheral leukocyte levels increased transiently (overshoot), and at 2 h after the completion of the treatment, they reached 193.4% of the baseline value. Since LCP is capable of reducing the peripheral blood leukocyte count over a short period of time, its impact on peripheral blood is great. In addition, in view of the overshoot phenomenon and the appearance of immature granulocytes, the LCP may affect not only the peripheral blood, but also the bone marrow pool, the marginal pool, and the leukocytes present in the tissues.     

Associations of Baseline and Changes in Leukocyte Counts with Incident Cardiovascular Events: The Dongfeng-Tongji Cohort Study

Aim: The aim of the present study was to investigate the associations of baseline and longitudinal changes in leukocyte counts with incident cardiovascular disease (CVD). Methods: We conducted a prospective study to investigate the associations of baseline and 5-year changes in leukocyte counts with incident CVD and its subtypes in middle-aged and elderly Chinese. We estimated the hazard ratios (HRs) and 95% confidence intervals (CIs) for CVD using the Cox proportional-hazards models. Results: In the analyses of baseline total leukocyte count of 26,655 participants, compared with the lowest quartile (＜4.71×10⁹/L), participants in the fourth quartile (＞6.70×10⁹/L) had 11% higher risk for CVD. Consistent with total leukocyte count, neutrophil count also exhibited a significant positive association with the risk of CVD. In the analyses of 5-year changes in total leukocyte count of 11,594 participants, the changes in leukocyte count were categorized into three groups, i.e., the decreased group (＜25%), stable group (25%–75%), and increased group (＞75%). Compared with participants in the stable group (−1.18 to 0.44×10⁹/L), participants in the increased group (＞0.44×10⁹/L) had 14% higher risk for CVD. We also observed significant positive associations of the changes in neutrophil and monocyte counts with the risk of CVD. Furthermore, the total leukocyte count in the second or third tertile at the first follow-up with a 5-year increase was related to higher CVD risk. Conclusion: High baseline total leukocyte count and a 5-year increase in total leukocyte count were related to higher CVD risk.     

Correlation Between Polymorphonuclear Leukocyte Counts and Levels of Tumor Necrosis Factor-a in Pleural Fluid of Patients with Parapneumonic Effusion

High concentrations of tumor necrosis factor-a (TNF), as well as excess of polymorphonuclear leukocytes (PMNs), are present in pleural fluid of patients with complicated parapneumonic effusion (CPPE). Several studies have shown that PMNs are capable of secreting TNF. However, the correlation between levels of TNF and PMN counts in pleural fluid of patients with parapneumonic effusion has not been previously evaluated. This study was undertaken to evaluate this correlation. Using a commercially available high sensitivity ELISA kit, levels of TNF were measured in pleural fluid of patients with UCPPE (n = 22), and CPPE (n = 14), and were compared with PMN count in pleural fluid of these patients. The mean ± SD values of pleural fluid TNF and PMN count in the UCPPE group the group were 10.15 ± 6.48 pg/mL and 3452 ± 2878 cells/mm3, respectively, and in the CPPE group the values were 55.51 ± 29.49 pg/mL and 25261 ± 11733 cells/mm3, respectively. Levels of pleural fluid TNF and PMN counts in the CPPE group were significantly higher than in the UCPPE group (p <0.0001). A significant correlation was found between levels of pleural fluid TNF and PMN counts in the CPPE group (r = 0.57, p = 0.03) and also in the UCPPE group (r = 0.44, p = 0.04). The results of this study indicate that in pleural fluid of patients with UCPPE or CPPE, levels of TNF correlate positively with PMN counts, and PMNs might be an important source of TNF production in pleural fluid of these patients, particularly in those with CPPE.     

Total Leukocyte Counts and the Requirement of Dose Reduction due to Cytopenias as Prognostic Indicators Affecting Response to Imatinib in Chronic Myeloid Leukemia

Imatinib is a tyrosine kinase inhibitor and is considered the first line of non stem cell transplantation treatment for patients diagnosed with CML. We evaluated the response rates and adverse reactions to Imatinib in our patients and tried to identify factors which affected the response to Imatinib. Eighty-four patients were diagnosed on the basis of clinical and haematological variables with confirmation by FISH, detecting Philadelphia chromosome or bcr-abl translocation and were then started on oral capsule Imatinib. A complete haematological response was seen in 78.04% patients, while complete cytogenetic response (CCR) was seen in 12.2% of patients and major cytogenetic response (MCR) was seen in 64.63% of patients. It was found that that a greater total leukocyte count (TLC) on presentation had a negative correlation with cytogenetic response. Cytopenias were seen in 36 patients (43.82%). 34.9% of patients having CCR/MCR required dose reduction while 73.6% of patients not achieving CCR/MCR required dose reduction. This was a significant difference, confirmed on statistical analysis (P < 0.05; P = 0.019), establishing the negative prognostic value of dose reduction due to cytopenias.     

白细胞计数和血小板计数联合评分对急性ST段抬高型心肌梗死患者急诊经皮冠状动脉介入治疗术后在院死亡危险评估的研究

目的：探讨白细胞计数和血小板计数联合评分(COL-P评分)对急性ST段抬高型心肌梗死（STEMI）患者急诊经皮冠状动脉介入治疗（PCI）术后在院死亡危险评估的效果。方法：回顾性研究我院2009-11至2013-08的STEMI住院急诊行PCI术的患者共660例,其中生存者572例,死亡者88例。依不同COL-P评分进行分组（COL-P 0分组、COL-P 1分组、COL-P 2分组）统计学分析。结果：660例急诊行PCI术的STEMI患者住院期间死亡者88例。死亡者白细胞计数高于生存者白细胞计数,两者比较差异有统计学意义（P<0.001）;死亡者血小板计数低于生存者血小板计数,两者比较差异有统计学意义（P<0.01）。Logistic回归显示,COL-P评分[COL-P（1 vs 0）,OR 4.346,95% CI：2.134-8.850,P<0.001; COL-P(2 vs 0), OR 10.126,95% CI：4.061-25.250,P<0.001]为STEMI患者急诊PCI术后在院死亡的独立影响因素。COL-P 0分组、COL-P 1分组和COL-P 2分组在院期间死亡率分别为4.9%、15.4%和43.1%,三组比较差异有统计学意义（P<0.001）。结论：COL-P评分是STEMI患者急诊PCI术后住院期间死亡率危险评估的良好评价工具,但对长期死亡率的评估能力还有待进一步研究验证。     

Establishment of a Small Animal Model for Human Helicobacter pylori Infection Using Germ-Free Mouse

Objective : To understand why oral inoculation of Helicobacter pylori resulted in continuous colonization of the stomach in germ-free athymic mice, but only temporary colonization in mice that were not germ-free. Methods : We inoculated germ-free and "not-germ-free" euthymic mice with H. pylori and studied the resulting colonization of the stomach, comparing it against the germ-free athymic mouse model. In addition, we inves-tigated Lactobacillus in the above-described three mouse groups. Results : H. pylori were detected in all germ-free athymic mice and all germ-free euthymic mice continuously. However, in all euthymic mice that were not germ-free, H. pylori was detected only tem-porarily after inoculation. Lactobacilli were detected only in the not-germ-free mouse group. The number of H. pylori in the germ-free euthymic mice was signifi-cantly lower than in the germ-free athymic mice during the period of this study after inoculation. Conclusions : We therefore suggest that the growth of H. pylori may be suppressed by the immunological sytem and eradi-cated by Lactobacilli previously inhabiting the stomach.     

Neonatally administered tert-octylphenol affects onset of puberty and reproductive development in female rats

There now is evidence that many of the synthetic chemicals released into the environment can impact on the function of the endocrine system of many organisms. One group of chemicals, the alkylphenols, used in paints, pesticides, herbicides, detergents, and plastics, has been found to have the ability to bind estrogen receptors. This estrogenic property makes these compounds potentially hazardous to the developing reproductive system and neuroendocrine brain. In this study we determined the effects of exposure to the environmental toxins 4-nonylphenol (NP) and 4-tert-octylphenol (OP) and to synthetic estrogen diethylstilbesterol (DES) during the early postnatal period (d 0–10) on the development of reproductive function. The day of vaginal opening, ovulation, prepubertal LH levels, LH response to estradiol, estrous cyclicity, and ovarian histology were determined. In the OP- and DES-treated groups, the vaginal opening was observed to have occurred several days prior to that of the control group. The NP-treated group showed vaginal opening at ages similar to those of the control group. Treatment with OP prevented ovulation in a significant number of animals, as well as in all animals treated with DES, whereas the control and NP-treated animals ovulated normally. Animals treated with DES and OP had significantly lower ovarian weights and higher uterine weights than either control animals or NP-treated animals. Higher basal LH levels, as well as the absence of the prepubertal LH surge, were observed in both DES- and OP-treated animals. A significant number of OP-treated animals showed no LH response to the estradiol-17β challenge. NP-treated animals responded positively to the estradiol-17β challenge. Persistent estrus was also apparent in both OP- and DES-treated animals. Upon histological examination, the ovaries in OP-treated animals were found to have a decreased number of corpora lutea and an increased number of preantral and atretic follicles. These data suggest that exposure to OP during the critical period of sexual brain differentiation affects the onset of puberty and reproductive development.     

Leukocyte reserves of newborn infants

Summary and Conclusions Leukocyte reserves of seven newborn infants have been estimated during therapeutic exchange transfusion, which had effected a leukopheresis. The results will be summarized at the end of the second paper [24], together with data on the restoration of new leukocyte circulating levels after the completion of the exchange transfusion. It then will be possible to conclude whether thetotal leukocyte storage pools had been estimated during ET, or whether additional undetected cell supplies had existed.

---

Zusammenfassung und Schlußfolgerung Durch eine therapeutische Austauschtransfusion wurde bei sieben Neugeborenen eine Leukophorese induziert, die eine Bestimmung der Leukozytenreserven dieser Kinder erlaubt. Die Ergebnisse werden mit den Beobachtungen der Wiederherstellung neuer Leukozytenspiegel nach Beendigung der Austauschtransfusion am Ende der zweiten Arbeit [24] zusammengefaßt. Erst dann wird beurteilt werden können, ob durch die Austauschtransfusion diegesamten Leukozytenvorräte erfaßt werden konnten, oder ob noch weitere Zellreserven vorhanden waren.

---

---

Supported by a grant from the Deutsche Forschungsgemeinschaft.

---

Presented at the XIIIth International Congress of Hematology, Munich 1970.     

Leukocyte reserves of newborn infants

Summary and Conclusions Leukocyte reserves of 7 newborn infants had previously been calculated during therapeutic exchange transfusion (ET) [10], and the post-ET restoration of new blood circulating levels have been observed in the present study:Reserves of segmented neutrophils about equal in size to the circulating pools before exchange transfusion, were present during the procedure in all infants. The supply was not exhausted in 5 full-term newborns, but the 2 premature infants showed a considerble delay in the restoration of new circulating levels post-ET.Blood lymphocyte reserves were availble during ET in 6 infants. They were exhausted during the procedure or after an additional limited release of cells post-ET. The subsequent increase in the circulating levels was not completed in most cases by 96 hours.Eosinophil reserves during ET were 2 to 3 times larger than the initial circulating pool in all infants. Since post-ET the establishment of a new circulating level was delayed beyond the eosinophil maturation time, the calculated values may represent the total size of the eosinophil reserves of these infants.Monocyte reserves were present in 6 infants. The post-ET restoration of a new circulating level occured in 5 infants within 3 to 6 hours, which is compatible with the short maturation time and the small cell reserves of monocytes in adults.

---

Zusammenfassung und Schlußfolgerung Die Leukozytenspeicher von 7 Neugeborenen waren während einer therapeutischen Austauschtransfusion in der vorausgegangenen Arbeit berechnet worden [10]; die Wiederherstellung neuer Leukozyten-Blutspiegel nach Beendigung des Austausches wurde in der vorliegenden Arbeit gemessen:Während des Austausches waren die Speicher neutrophiler Segmentkerniger bei allen Neugeborenen etwa in Höhe des ursprünglichen Blutspiegels nachweisbar. Nach dem Austausch erwiesen sich die Speicher bei 5 Reifgeborenen als nicht ausgeschöpft; dagegen zeigten die 2 Frühgeborenen eine deutlich verzögerte Wiederherstellung des Neutrophilenblutspiegels.Blutlymphozytenspeicher-bei 6 Probanden während des Austausches nachgewiesen-waren entweder am Ende des Austausches oder nach einer zusätzlichen, begrenzten Ausschüttung von Zellen in den ersten Stunden nach dem Austausch ausgeschöpft. Der Wiederanstieg der Blutlymphozyten war in den meisten Fällen nach 96 Stunden nicht abgeschlossen.Eosinophilenspeicher in doppelter bis dreifacher Höhe des ursprünglichen Blutspiegels bestanden während des Austausches bei allen Neugeborenen. Da ein neuer Eosinophilenblutspiegel nicht vor Ablauf der Eosinophilenausreifungszeit wiederhergestellt wurde, dürfte die Gesamtgröße der Eosinophilenspeicher der Neugeborenen erfaßt worden sein.Monozytenspeicher fanden sich bei 6 Kindern. Die Wiederherstellung eines neuen Blutspiegels im Anschluß an die Austauschtransfusion bei 5 Neugeborenen innerhalb von 3 bis 6 Stunden stimmt mit der kurzen Ausreifungszeit und den kleinen Speichern der Monozyten bei Erwachsenen überein.

---

---

Supported by a grant from the Deutsche Forschungsgemeinschaft.

---

Presented at the XIIIth International Congress of Hematology, Munich 1970.