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Related Concept Videos

Muscles of the Shoulder01:23

Muscles of the Shoulder

The muscles surrounding the shoulder girdle, including the clavicle and scapula, primarily stabilize the scapula. This stable base allows other muscles to move the humerus effectively. Scapular movements often mirror those of the humerus and extend its range of motion. For instance, raising the arm above the head would not be feasible without simultaneous upward rotation of the scapula.
Anterior Thoracic Muscles
The anterior thoracic muscles include the serratus anterior, subclavius, and...
Muscles that Move the Arm01:31

Muscles that Move the Arm

Nine muscles are involved in arm movements. Two of these, the pectoralis major and latissimus dorsi, originate from the axial skeleton and are called axial muscles. The other seven originate from the scapula and are called the scapular muscles.
The pectoralis major has two origins. Its clavicular head originates on the medial half of the clavicle. In contrast, the sternocostal head originates on the costal cartilages of ribs 1-6, the sternum, and the aponeurosis of the external oblique of the...

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Assessing the Immunogenicity of Synthetic RNA Using Blood Cells.

Mouldy Sioud1

  • 1Division of Cancer Medicine, Department of Cancer Immunology, Institute for Cancer Research, Oslo University Hospital Radiumhospitalet, Oslo, Norway. mosioud@medisin.uio.no.

Methods in Molecular Biology (Clifton, N.J.)
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Summary

Evaluating the immunostimulatory potency of therapeutic RNA is crucial. This chapter details methods using peripheral blood mononuclear cells (PBMCs) to assess RNA-induced innate immune activation and cytokine production.

Keywords:
Blood cellsInflammatory cytokinesInnate immunityPattern recognition receptorsRNAType I interferonsmRNA

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Area of Science:

  • Immunology
  • Molecular Biology
  • Pharmacology

Background:

  • The innate immune system utilizes pattern recognition receptors (PRRs) to detect foreign nucleic acids, initiating signaling cascades that result in pro-inflammatory and interferon responses.
  • While vital for antiviral defense, these immune reactions can compromise RNA-based therapeutics, causing immunogenicity, toxicity, and reduced efficacy.
  • Assessing the immunogenicity of therapeutic RNAs is therefore a critical step in their development.

Purpose of the Study:

  • To provide detailed protocols for evaluating the immunostimulatory potential of exogenous RNA.
  • To establish methods for assessing innate immune activation in response to RNA-based therapeutics.
  • To offer a framework for the safety and efficacy evaluation of RNA therapeutics.

Main Methods:

  • Isolation of peripheral blood mononuclear cells (PBMCs) from human blood.
  • Stimulation of PBMCs or whole blood with synthetic or in vitro-transcribed mRNA.
  • Quantification of cytokine production using enzyme-linked immunosorbent assays (ELISA).

Main Results:

  • Established protocols for measuring RNA-induced immune responses in vitro.
  • Demonstrated the utility of PBMCs and whole blood as models for innate immune activation.
  • Provided a method for quantifying cytokine release as a marker of immunostimulatory potency.

Conclusions:

  • The presented protocols enable robust assessment of RNA immunostimulatory activity.
  • These methods are essential for the preclinical evaluation of RNA-based therapeutics.
  • Accurate assessment of immunogenicity is key to developing safe and effective RNA medicines.