High Concentrations Of Leukocytes Research Articles

Prolonged administration of oral phenylbutazone and firocoxib in horses has no impact on selected cytokine and growth factor concentrations in platelet-rich plasma and autologous protein solution.

To determine the effects of prolonged administration of the oral NSAIDs phenylbutazone and firocoxib on concentrations of cytokines and growth factors in platelet-rich plasma (PRP) and autologous protein solution (APS). 6 adult University owned horses. Horses were randomized to receive phenylbutazone (1 g, orally, q 12 h) or firocoxib (57 mg, orally, q 24 h) for 6 days. Blood was obtained and processed for APS (Pro-Stride) and PRP (Restigen) before the administration of NSAIDs and at 7 days (1 day following cessation of NSAIDs). Horses underwent a two-week washout period, during which blood was obtained at 14 days and 21 days. The protocol was repeated with a crossover design. PRP and APS were analyzed for concentrations of platelets, leukocytes, and several cytokines (IL-1β, IL-10, IL-6, IL-8, and tumor necrosis factor-α) and growth factors (PDGF, FGF-2, and TGF-β1) using immunoassays. Plasma was evaluated for drug concentrations. No significant differences existed in concentrations of growth factors and cytokines before or after prolonged administration of NSAIDs. There were significant differences in concentrations of leukocytes and platelets in PRP compared to APS, with higher concentrations of leukocytes at the day 7 time point (T) in APS (phenylbutazone) and in concentrations of platelets in APS at T0 (firocoxib) and in APS at T7 (phenylbutazone). Veterinarians can recommend the administration of these oral NSAIDs prior to obtaining blood for PRP and APS provided a single-day washout period is instituted.

Platelet-Rich Plasma Injections for Chronic Tendinopathies in the Upper Extremities


 Findings from evidence syntheses describing the comparative clinical effectiveness of platelet-rich plasma injection (PRPi) in the treatment of chronic tendinopathies of the upper extremities were variable.
 Whereas findings describing function showed no clear demonstration of effect, some findings describing pain suggested there may be potential for a benefit of PRPi with higher concentrations of leukocytes or longer durations of follow-up.
 Lack of a clear demonstration of the comparative clinical effectiveness of PRPi in chronic tendinopathies of the upper extremities does not currently support decision-making in favour of its use.

Autologous platelet concentrates in regenerative endodontic treatment

Introduction: Autologous platelet concentrates, have recently emerged as a possible tool to improve regenerative procedures in the medical field. Objective: To analyze the literature on autologous platelet concentrates in regenerative endodontics, such as: Platelet Rich Plasma, Platelet Rich Fibrin, PRP/PRF Vs Blood Clot as a scaffold in regenerative endodontics. Methodology: Using the keywords Regenerative Endodontic Treatment, Autologous Platelet Concentrates, Immature Teeth, Platelet-rich Fibrin, Platelet-rich Plasma, Revitalization Endodontics, the MEDLINE/PubMed and ScienceDirect databases were searched, with emphasis on the last 5 years. It was evaluated with the PRISMA and AMSTAR-2 guidelines. Results: Platelet-rich plasma releases growth factors during the first hours after placement and induction of mesenchymal stem cells. Platelet Rich Fibrin is a second-generation platelet concentrate that releases growth factors more slowly, forming a network that traps leukocytes and platelets, liquid platelet rich fibrin has a higher concentration of leukocytes, which enhances immune defense in LPS-induced inflammation. Compared to blood clot, PRP and PRF show improved healing results, reducing infection and inflammation. Conclusion: The use of autologous platelet concentrates presents a more controlled scaffold in regenerative endodontics, with successful results, such as periapical healing, root thickening and apical closure.

Allogeneic transplant procurement in the times of COVID-19: Quality report from the central European cryopreservation site

BackgroundBecause of limitations of transportation imposed by the COVID-19 pandemic, current recommendation calls for cryopreservation of allogeneic stem cell transplants before patient conditioning. A single cell therapy laboratory was selected to function as the central cryopreservation hub for all European registry donor transplants intended for the Australian-Pacific region. We examined properties of these transplants to ascertain how quality is maintained.MethodsWe analyzed 100 pandemic-related allogeneic mobilized blood-derived stem cell apheresis products generated at 30 collection sites throughout Europe, shipped to and cryopreserved at our center between April and November of 2020. Products were shipped in the cool, subsequently frozen with DMSO as cryoprotectant. Irrespective of origin, all products were frozen within the prescribed shelf-life of 72 h.ResultsPrior to cryopreservation, viable stem cell and leukocyte count according to the collection site and our reference laboratory were highly concordant (r2 = 0.96 and 0.93, respectively) and viability was > 90% in all instances. Median nominal post-thaw recovery of viable CD34+ cells was 42%. Weakly associated with poorer CD34+ cell recovery was higher leukocyte concentration, but not time lag between apheresis or addition of cryopreservant, respectively, and start of freezing. The correlation between pre- and post-thaw CD34+ cell dose was high (r2 = 0.85), hence predictable. Neutrophil and platelet engraftment were prompt with no evidence of dose dependency within the range of administered cell doses (1.31–15.56 × 106 CD34+ cells/kg).ConclusionsGeneral cryopreservation of allogeneic stem cell transplants is feasible. While more than half of the CD34+ cell content is lost, the remaining stem cells ensure timely engraftment.

Quality comparison between two different types of platelet-rich plasma for knee osteoarthritis.

Introduction: Knee osteoarthritis (KOA), the most common form of osteoarthritis (OA) is a considerable health concern worldwide. Platelet-rich plasma (PRP) is a common therapeutic option for KOA. Different types of PRPs have varying efficacies. However, a comparative analysis of the qualities of these PRPs is lacking. Methods: Two types of PRPs, including autologous protein solution (APS), and leukocyte-poor PRP (LP-PRP) along with whole blood (WB) and platelet-poor plasma (PPP) were characterized for platelet content, leukocyte content, and composition in 10 healthy volunteers (HV) (the controlled laboratory study) and 16 KOA patients (a retrospective observational study). Additionally, the levels of the platelet-derived growth factor (PDGF)-BB, and different cytokines were estimated in HV. Results: In HV, the concentrations of platelets and leukocytes, levels of different cytokines, including interleukin 1 receptor antagonist (IL-1Ra), soluble TNF receptor type II (sTNF-RII), and IL-1β, and the ratio of IL-1Ra/IL-1β were significantly higher in APS, whereas the PDGF-BB was higher in LP-PRP than APS. In KOA patients, a higher concentration of platelets was observed in LP-PRP, and a higher concentration of leukocytes was observed in APS than LP-PRP. Following the PAW classification system, LP-PRP was classified as P2-B type in HV (51.3 × 104/μl) and KOA (53.4 × 104/μl), whereas APS was classified as P3-A type in HV (110.1 × 104/μl) and P2-A type in KOA (29.0 × 104/μl). In a retrospective observational study, the KOA patients who underwent APS injection had a higher incidence of arthralgia, and this arthralgia lasted for a longer time than LP-PRP injection in the same individual. Discussion: The quality of the two PRPs differed distinctively depending on their preparation methods, which might affect their clinical efficacies and adverse events. Therefore, the characterization of these parameters should be prioritized while choosing PRP.

Total and Differential Leukocyte Counts in Relation to Incidence of Diabetes Mellitus: A Prospective Population-Based Cohort Study.

ObjectiveHigh concentrations of leukocytes in blood have been associated with diabetes mellitus. This prospective study aimed to explore whether total and differential leukocyte counts are associated with incidence of diabetes. A missense variant R262W in the SH2B3 (SH2B adaptor protein 3) gene, coding for a protein that negatively regulates hematopoietic cell proliferation, was also studied in relation to incidence of diabetes.Methods and ResultsLeukocyte count and its subtypes (neutrophils, lymphocytes and mixed cells) were analyzed in 26,667 men and women, 45–73 years old, from the population-based Malmö Diet and Cancer study. Information about the R262W polymorphism (rs3184504) in SH2B3 was genotyped in 24,489 subjects. Incidence of diabetes was studied during a mean follow-up of 14 years. Cox proportional hazards regression was used to examine incidence of diabetes by total and differential leukocyte counts. Mendelian randomization analysis using R262W as an instrumental variable was performed with two-stage least squares regression. A total of 2,946 subjects developed diabetes during the follow-up period. After taking several possible confounders into account, concentrations of total leukocyte count, neutrophils and lymphocytes were all significantly associated with incidence of diabetes. The adjusted hazard ratios (95% confidence interval; quartile 4 vs quartile 1) were 1.37 (1.22–1.53) for total leukocytes, 1.33 (1.19–1.49) for neutrophils and 1.29 (1.15–1.44) for lymphocytes. The R262W polymorphism was strongly associated with leukocytes (0.11x109 cells/l per T allele, p = 1.14 x10-12), lymphocytes (p = 4.3 x10-16), neutrophils (p = 8.0 x10-6) and mixed cells (p = 3.0 x10-6). However, there was no significant association between R262W and fasting glucose, HbA1c or incidence of diabetes.ConclusionsConcentrations of total leukocytes, neutrophils and lymphocytes are associated with incidence of diabetes. However, the lack of association with the R262W polymorphism suggests that the associations may not be causal, although limitations in statistical power and balancing pleiotropic effects cannot be excluded.

Goat cathelicidin-2 is secreted by blood leukocytes regardless of lipopolysaccharide stimulation.

It has been reported that goat cathelicidin-2, an antimicrobial peptide, localizes in leukocytes and is present in milk. Here, we examined whether cathelicidin-2 is secreted by leukocytes. Different concentrations (10(5)-10(8) cells/mL) of blood leukocytes were cultured for 0-48 h with or without lipopolysaccharide (LPS). After culture, the concentrations of cathelicidin-2 in the conditioned media were measured. Blood was collected from male goats 0-24 h after the intravenous injection of Escherichia coli O111:B4 LPS. The plasma cathelicidin-2 concentrations were determined and the blood leukocytes immunostained with anti-cathelicidin-2 antibody to calculate the proportion of cathelicidin-2-positive cells in the total leukocytes. When higher concentrations of leukocytes were cultured, the cathelicidin-2 concentrations in the media increased significantly, whereas the addition of LPS to the media caused no further increase. The plasma cathelicidin-2 concentrations did not increase with time after LPS infusion. The proportion of cathelicidin-2-positive cells in the total leukocytes was significantly reduced 1 h after LPS injection compared with that at 0 h, but increased again at 6 h and thereafter. These results suggest that cathlicidin-2 is secreted by leukocytes even without LPS stimulation, whereas LPS may be required for cathelicidin-2-containing leukocytes to be recruited from the blood to tissues showing inflammation.

High leukocyte count and interleukin-10 predict high on-treatment-platelet-reactivity in patients treated with clopidogrel

According to recent trials, a significant number of patients do not have a completely effective response to clopidogrel. The aim of the study was to evaluate the rate of clopidogrel resistance in the context of important clinical characteristics and to specifically determine the relation between clopidogrel efficacy and biomarkers of inflammation. Consecutive non-selected patients following PCI were enrolled into the study. All patients received a loading dose of 600mg of clopidogrel. The effect of clopidogrel was assessed using the VerifyNow assay 24h after clopidogrel administration, clopidogrel resistance was defined as PRU≥240. At the same time, standard parameters of biochemistry and hematology, the concentration of anti-inflammatory cytokine interleukin-10 and of soluble CD40 ligand, were measured. 378 patients were enrolled. 243 (64.3%) patients were responders (R) and 135 patients (35.7%) were non-responders (NR). Non-responders were older (R 65.7±13.3, NR 69.8±11.5, P<0.05), had a higher prevalence of diabetes (R 26.3%, NR 38.5%, P<0.05), were more often on mechanical ventilation (R 0.8%, NR 4.4%, P<0.05). The leukocyte count (R 9.8±3.5, NR 11.7±12.8, P<0.05), and concentration of IL-10 (R 3.1pg/ml, NR 5.7pg/ml, P<0.05) was higher among non-responders. The concentration of CD40L was not significantly different between the groups. In a multivariate logistic regression, older age, higher weight, female gender, mechanical ventilation, and a higher concentration of leukocytes and IL-10 were associated with an increased risk for being a non-responder. Older, obese patients, especially women had a higher risk of high on-treatment-platelet-reactivity. Higher concentrations of leukocytes and interleukin-10 were also an important factor associated with the risk of low clopidogrel responsiveness.

Carcinoembryonic antigen and neutrophils in healthy smokers

We measured the levels of carcinoembryonic antigen (CEA) and the concentrations of neutrophils in serum from healthy smokers. The smokers (n = 1396) had more leukocytes, neutrophils, and lymphocytes than did non-smokers (n = 1123) (leukocytes: 5853 +/- 1627/mm3 vs 4564 +/- 1118/mm3, p < 0.0001; neutrophils: 3533 +/- 1280/mm3 vs 2718 +/- 883/mm3, p < 0.0001; lymphocytes; 1843 +/- 502/mm3 vs 1513 +/- 406/mm3, p < 0.0001). Subjects in whom the levels of CEA was greater than 5.0 ng/ml had significantly higher concentrations of leukocytes, neutrophils and lymphocytes than did those in whom it was 5.0 ng/ml or less (p < 0.0001). Weak correlations were found between serum CEA levels and both the duration of smoking and the Brinkman index. The CEA levels also correlated with daily cigarette consumption (rho = 0.53, p < 0.0001). The concentration of neutrophils in subjects who smoked no more than 20 cigarettes per day or had a CEA level greater than 5.0 ng/ml, or both, was higher than that in subjects who smoked more than 20 cigarettes per day or had a CEA level less than or equal to 5.0 ng/ml, or both (4097 +/- 1325/mm3 vs 3791 +/- 1278/mm3, p < 0.05). These results suggest that in healthy smokers, high serum CEA levels are related to high neutrophil levels.

Age and environment affect constitutive immune function in Red Knots (Calidris canutus)

We studied subspecies, age and environmental effects on constitutive immune function (natural antibody and complement titres, haptoglobin activity and leukocyte concentrations) in Red Knots (Calidris canutus). We compared C. c. islandica and C. c. canutus in the Wadden Sea and found no difference in immune function between subspecies. However, C. c. canutus on their wintering grounds in Banc d’Arguin had higher natural antibody and lower complement levels than C. c. canutus or C. c. islandica in the Wadden Sea. This suggests that immune function is determined more by the surrounding environment than by subspecies. We also compared age classes in the Wadden Sea and found that first year birds had significantly lower natural antibody levels than adults, but that second year birds no longer differed from adults. Finally, we examined the interaction of age and environment in Banc d’Arguin. We found that first year birds (but not adults) in a low quality habitat had higher leukocyte concentrations than first year birds or adults in a high quality habitat. Differences in available resources and defence needs between environments, and differences among individuals differentially distributed between sites, are likely important contributors to the variation in immune function we report. Future studies, which examine these factors on wild birds, will be important for our understanding of how animals function in their natural environment. (220).

Cost of parasitism and host immune defence in the sand martin Riparia riparia: a role for parent-offspring conflict?

Parent-offspring conflict may arise because the lifetime reproductive success of the parent is more influenced by its life span than by reproductive success during a particular reproductive event, while the fitness of an offspring depends firstly on its own survival as a juvenile and only subsequently on its own reproductive success. The naive immune system of young animals may allow offspring to be much more affected by parasites than their parents, and thus cause an initial asymmetry in a potential parent-offspring conflict. We investigated this type of conflict by assessing the health status and the immune response of parent and offspring sand martins Riparia riparia infested with manipulated loads of ticks Ixodes lividus (nests either treated with pyrethrin, water, or just visited). The prevalence and the intensity of tick infestations differed among treatments, with low tick loads in nests with the pyrethrin treatment. Ticks reduced the reproductive success of the host and increased offspring wing length. Broods with ticks had higher leukocyte concentrations and concentrations of immunoglobulins. The concentration of immunoglobulins in nestlings was negatively related to brood size and nestling tarsus length. Nestlings receiving the control treatments had a positive association between wing length and the concentration of immunoglobulins and a negative association between tarsus length and immunoglobulins. In contrast, adult sand martins did not respond to the parasite treatment in terms of immune response. Hence, the naive immune system of nestlings may be the crucial factor causing the parent-offspring conflict over costs of parasitism to be resolved to the advantage of parents that may sacrifice nestlings in heavily parasitized nests.

Parasites, Immunology of Hosts, and Host Sexual Selection

Parasite-mediated sexual selection is reviewed with special emphasis on the bird literature. Choosy females may benefit from choosing parasite-free mates if such males provide better parental care, do not transmit contagious parasites, or provide resistance genes to offspring. There is evidence in support of each of these mechanisms. The immunocompetence handicap hypothesis posits that secondary sexual characters reliably reveal the ability of males to resist parasites due to the immunosuppressive effects of testosterone and other biochemicals. Several aspects of these negative feedback mechanisms are supported by laboratory studies, but evidence from free-living animals is almost completely absent. Corticosterone rather than testosterone may potentially mediate the immunocompetence handicap mechanism. A simple version of the immunocompetence handicap is developed suggesting that body condition of male hosts is a sufficient mediator of the handicap mechanism of reliable sexual signaling. Sexual selection appears to be more intense in sexually dichromatic bird species, and comparative studies using pairwise comparisons of closely related taxa reveal that sexually dichromatic bird species have larger spleens, larger bursa of Fabricius, and higher concentrations of leukocytes than monochromatic species. Parasite-mediated sexual selection is proposed to affect parasite biology by increasing (1) the variance-to-mean ratio in parasite abundance, (2) variance in the intensity of natural selection affecting hosts, and (3) speciation rates among parasites exploiting hosts subject to intense sexual selection as compared to those subject to less intense selection.

White blood cells in semen affect hyperactivation but not sperm membrane integrity in the head and tail regions

The presence of high numbers of peroxidase-positive PML in ejaculated semen significantly reduced sperm HA, an important step leading to sperm capacitation. Sperm membranes at both the head and tail regions, as assessed by the hypo-osmotic viability parameter and the hypo-osmotic sperm swelling test, respectively, were not affected by peroxidase-containing leukocytes. Sperm motility was not affected, but sperm curvilinear and straight line velocity parameters were reduced in the presence of high concentrations of leukocytes in the ejaculate. The results suggested that the effect of leukocytes on sperm was through a reduction in sperm hyperactive motility but not through alterations in the sperm head and tail membranes.

Leukocyte distribution in the pseudopregnant mouse uterus.

Pseudopregnancy, induced by mating females with vasectomized males, is a frequently used model for studying pregnancy-like uterine changes in the absence of an embryo. Leukocytes make a significant contribution to uterine cellularity during pregnancy. The present study was designed to determine whether changes in numbers and distribution of leukocytes in the uterus during pseudopregnancy and following intraluminal injection of a deciduogenic stimulus parallel changes observed during the first eight days of pregnancy. Common leukocyte antigen positive (CLA+) cells, macrophages (F4/80+ cells), and granulocytes were assessed between days 1 and 8 of pseudopregnancy using qualitative and quantitative immunohistochemistry. High numbers of CLA+ leukocytes were present on days 1 and 2. Those were comprised primarily by macrophages, polymorphonuclear leukocytes, and eosinophils. High concentrations of leukocytes were detected in the endometrium, and some granulocytes were observed migrating through the luminal epithelium. Leukocytes, principally macrophages, were reduced in number and were distributed throughout the endometrium on days 3 and 4. Introduction of oil to the uterine lumen on day 4 stimulated primary decidualization. Decidual cells were CLA- and F4/80-, and, as decidualization proceeded, CLA+ and F4/80+ cells decreased in number in the anti-mesometrial uterus and were detected primarily in the deep endometrium. Later, a secondary decidualization zone developed in the mesometrial aspect of the uterus. Unlike the initial decidual reaction, which was relatively free of leukocytes, the secondary decidual zone contained very high numbers of CLA+ and F4/80+ cells. The uninjected uterine horn remained relatively unchanged from days 3 through 8.(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of lipid and leukocytes on the haemoglobin determination by Coulter Counter S Plus III, Technicon H 6000, Technicon H 1, LK 540, Reflotron and Hemocap

The influence of lipid and high concentrations of leukocytes on the haemoglobin determination by six different instruments was examined. The increase in the determination of haemoglobin concentration with elevated amounts of lipid in the samples was greatest with the Coulter Counter S plus III and the Technicon H 1 followed by the LK 540 and the Technicon H 6000. There was no effect of lipid on the haemoglobin determination using the Reflotron. High concentrations of leukocytes increased the haemoglobin determination most by Coulter Counter S plus III and Hemocap. There were no changes in the haemoglobin determinations by the LK 540 and the Technicon H 6000 when the samples contained increased amounts of leukocytes.

Preparation of Leukocyte‐Poor Platelet Concentrates from Buffy Coats

To study the influence of contaminating leukocytes on the storage conditions of platelet concentrates (PC), various amounts of leukocytes were added to identical PC. From 12 blood donations, 12 leukocyte-poor PC were prepared and pooled. Subsequently, the pool was divided into 12 identical PC. The plasma volume of the PC was 58.6 +/- 0.6 ml, the platelet concentration was 1.01 +/- 0.04 x 10(9)/ml (mean +/- SD) and the red cell contamination did not exceed 10(7) per PC. To 4 groups of 3 PC, pooled leukocytes were added from the same 12 blood donations. The leukocyte contamination for each group of 3 PC was 0.14 +/- 0.05, 1.96 +/- 0.09, 5.53 +/- 0.98 and 13.0 +/- 0.93 x 10(6)/ml (mean +/- SD) for groups I-IV, respectively. The PC were stored for 7 days at 22 degrees C in normal polyvinylchloride bags. A significant correlation was found between increasing concentrations of leukocytes in the PC and the drop in pH (r = -0.93), glucose consumption (r = -0.91), lactic acid production (r = 0.93) and release of lactate dehydrogenase (r = 0.92) during storage of the PC. The excretion of beta-thromboglobulin, depletion of platelet adenine nucleotides, decreased ability to incorporate 3H-adenosine into metabolic nucleotides and poor morphology of the platelets were also significantly correlated with an increased number of leukocytes in the PC. These data show that high concentrations of leukocytes in PC have a significant detrimental effect on the viability of platelets during storage at 22 degrees C. We conclude that for good storage conditions of PC, the upper limit of leukocytes per PC should not exceed 10(7).

Effect of High Concentrations of Leukocytes on Whole Blood Viscosity

Abstract Whole blood viscosity of patients with leukemic leukocytosis was compared to that of normals with a similar total packed cell volume (TPCV). Leukocytes in suspension were also studied and found to have a greater viscosity than erythrocytes. We found no relation between the magnitude of the leukocyte count and whole blood viscosity. The presence of anemia in the patients with leukemia prevented elevation of their TPCV. This, as well as the normal variability of blood viscosity obscured any increase that could have been produced by the greater viscosity of massed leukocytes.