Purification of Functional Platelet Mitochondria Using a Discontinuous Percoll Gradient

Léger, Jacob L; Pichaud, Nicolas; Boudreau, Luc H.

doi:10.1007/978-1-0716-1266-8_4

Cited by 4 publications

(4 citation statements)

References 19 publications

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“…When about 80% of Vero cells were rived by T. gondii RH or GT1 tachyzoites in vitro, the culture products were carefully scraped, fully mixed and broken, and purified using the density gradient centrifugation of Percoll (GE Healthcare, Uppsala, Sweden) ( 22 , 23 ). The obtained tachyzoites were washed thrice with centrifugation at 5 000 rpm for 3 min, counted by a hemacytometer, and adjusted to 2.0 × 10 6 parasites per milliliter for RH or GT1 strain using stroke-physiological saline solution (SPSS).…”

Section: Methodsmentioning

confidence: 99%

Immunization of BALB/c Mice with Killed Tachyzoites of Toxoplasma gondii against Acute Toxoplasmosis

Guo,

Tan,

et al. 2023

IJPA

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Background: Toxoplasma gondii with widespread distribution infects over one third of human populations in the world and can cause serious life-threatening diseases especially for the immunodeficient patients in acute toxoplasmosis. As the clinical pharmaceutical drugs with severe side effects for treatment and non-ideal extant vaccines for prevention, more work starves to be done for keeping advantages in the athletics. Methods: Aluminum adjuvant and hybrid formaldehyde-killed tachyzoites of T. gondii RH and GT1 isolates were prepared to intramuscularly immunize BALB/c mice for five times at 0, 3, 7, 14 and 21 days post first injection. The triggered humoral and cellular immune responses at two weeks post the last immunization and the survival times of infected mice were examined for the hybrid immunization scheme judgement. Results: The anti-RH and anti-GT1 specific antibodies were both increased at one week prior to challenge (P < 0.05), and the survival times of immunized mice (7.33 ± 0.71 d for RH, 7.22 ± 0.97 d for GT1) against acute toxoplasmosis were significantly prolonged by the immunizations performed in the study compared to blank control (6.67 ± 0.50 d for RH, 6.33 ± 0.71 d for GT1; P < 0.05), with the higher IFN-γ, IL-2 and IL-12p70 in sera, the elevated CD3e+CD4+ T and CD3e+CD8a+ T cells, and the enhanced lymphocyte proliferation in spleen (P < 0.05). Conclusion: The hybrid killed tachyzoites with aluminum adjuvant induced humoral and cellular immune responses of mice, and offered mildly protective efficacy against acute toxoplasmosis.

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Section: Methodsmentioning

confidence: 99%

Immunization of BALB/c Mice with Killed Tachyzoites of Toxoplasma gondii against Acute Toxoplasmosis

Guo,

Tan,

et al. 2023

IJPA

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“… A, BTransmission electron microscopy imaging of our mitochondria preparation and purification as previously reported (Léger et al , 2020, 2021). The black scale bars shown in the lower right of panels (A) and (B) represent sizes of 200 nm and 1 μm, respectively. …”

Section: Methodsmentioning

confidence: 99%

“…The pellet was then resuspended in IB and centrifuged at 13,000 g at 4°C for 10 min. Purification of platelet‐derived mitochondria is shown in Fig EV3A and B, as previously developed by our group (Léger et al , 2020, 2021).…”

Section: Methodsmentioning

confidence: 99%

Functional platelet‐derived mitochondria induce the release of human neutrophil microvesicles

et al. 2022

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Inflammation is an essential process of host defense against infections, illness, or tissue damage. Polymorphonuclear neutrophils (PMN) are among the first immune cells involved in acute inflammatory responses and are on the front line in the fight against bacterial infections. In the presence of bacterial fragments, PMN release inflammatory mediators, enzymes, and microvesicles in the extracellular milieu to recruit additional immune cells required to eliminate the pathogens. Recent evidence shows that platelets (PLTs), initially described for their role in coagulation, are involved in inflammatory responses. Furthermore, upon activation, PLT also release functional mitochondria (freeMitos) within their extracellular milieu. Mitochondria share characteristics with bacterial and mitochondrial damage‐associated molecular patterns, which are important contributors in sterile inflammation processes. Deep sequencing transcriptome analysis demonstrates that freeMitos increase the mitochondrial gene expression in PMN. However, freeMitos do not affect the mitochondrial‐dependent increase in oxygen consumption in PMN. Interestingly, freeMitos significantly induce the release of PMN‐derived microvesicles. This study provides new insight into the role of freeMitos in the context of sterile inflammation.

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“…Another possibility is that PRP directly reinforces the ribosomal and mitochondrial complement by sharing platelet organelles (presumably of better quality) with recipient tissue. Operating separately from cytokines or other growth factors discharged after activation, any platelet ribosomes [ 58 ] or platelet mitochondria [ 59 ] contributed by intraovarian PRP have yet to be specifically tracked.…”

Section: Subcellular Characteristics After Prpmentioning

confidence: 99%

Multichannel Recovery Potential with Activated Autologous Intraovarian Platelet-Rich Plasma and Its Derivatives

Sills

Wood

2023

Medicines

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Platelet-rich plasma (PRP) is an ‘orthobiologic’ with recognized roles in plastic surgery, musculoskeletal disorders, dentistry, dermatology, and more recently, ‘ovarian rejuvenation’. Intraovarian PRP involves a complex secretome discharged after platelet activation, comprising multiple cytokine mediators delivered surgically to older or inactive ovarian tissue. Loss of oocyte meiotic fidelity and impaired fertilization accompanying advanced maternal age are already managed by IVF, but only with eggs provided by younger donors. However, if the observed effect of rectifying embryo ploidy error can be proven beyond case reports and small series, activated PRP (or its condensed plasma cytokines) would deliver a welcome therapeutic disruption that is difficult to overstate. Because shortcomings in ovarian function are presently addressed mainly by pharmacological approaches (i.e., via recombinant gonadotropins, GnRH analogs, or luteal support), autologous PRP would represent an unusual departure from these interventions. Given the diversity of platelet cargo proteins, the target response of intraovarian PRP is probably not confined to oocytes or follicles. For example, PRP manipulates signal networks driving improved perfusion, HOX regulation, N-glycan post-translational modification, adjustment of voltage-gated ion channels, telomere stabilization, optimization of SIRT3, and ribosome and mitochondria recovery in older oocytes. While multichannel signals operating on various pathways are not unique to reproductive biology, in intraovarian PRP this feature has received little study and may help explain why its standardization has been difficult. Against this background, our report examines the research themes considered most likely to shape clinical practice.

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Purification of Functional Platelet Mitochondria Using a Discontinuous Percoll Gradient

Cited by 4 publications

References 19 publications

Immunization of BALB/c Mice with Killed Tachyzoites of Toxoplasma gondii against Acute Toxoplasmosis

Immunization of BALB/c Mice with Killed Tachyzoites of Toxoplasma gondii against Acute Toxoplasmosis

Functional platelet‐derived mitochondria induce the release of human neutrophil microvesicles

Multichannel Recovery Potential with Activated Autologous Intraovarian Platelet-Rich Plasma and Its Derivatives

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