Category Archives: Immunology

Gene Silencing Technology

Year 2018 welcomes the novel family of FDA approved drugs for inherited diseases. These drugs which are backed by a Nobel Prize and 2 decades of research have the ability to cure inherited diseases without actually editing the delicate genome. It is said that they are powerful enough to give a backseat to CRISPR based gene therapy. One such drug is “Patisiran” developed by biopharmaceutical company-Alnylam for treatment of genetic nerve damage. It works on RNA interference (RNAi) technology.

As we all know that DNA along with messenger RNA (mRNA) produces proteins via transcription and translation process. RNAi does the work of shooting down the messenger and erasing the message for protein production. Scientists believe that this will be greater method for treatment of inherited genetic or immune disorders than gene therapy or immunotherapy. Gene therapy involves cutting out the mutated gene region and replacing it with the new gene but this tampering may have certain risks like ethical regulations, unspecific gene region cut and unpredicted changes in gene silently leading to cancer activation. However RNAi ensures that no bad disease is developed and no bad protein is produced by shooting down the compilation of the message by mRNA.akk

RNAi not just works on inherited diseases but also on non-inherited diseases like stroke too. In Stroke, group of proteins get activated together to kill the brain cells. RNAi can stop such activation thus protecting the brain cells. Sometimes even good proteins can turn bad upon being faced by any pathogenic situation leading to a disease. As RNAi are reversible when compared to gene therapy, RNAi can be used to silence the proteins temporarily in such circumstances and restore them back to their normal functional state after the change in situation.

So the question arises. If function of RNAi is really that great and it was discovered long back in 1998, what’s stopping this RNAi or RNA technology developed drugs to enter the market? It seems that the production in this technology is easy but the real problem lies in targeting and delivering the therapy to the specified tissue in question. RNAi works by addition of small snippets of synthetic nucleotides which homes the RNAi, delivers it to the disease-causing RNA to silence it. The problem that arises is host immune system gets triggered upon the introduction of these nucleotide snippets in the body and the activation can lead to massive inflammation and even death. To avoid the immune system risk these snippets can be coated in nanoparticles but they will just end up in liver or kidney thus making the RNAi treatment difficult to reach brain, heart or lungs.

Patisiran was successful and approved by FDA for treatment of rare disease called hereditary transthyretin-mediated amyloidosis (hATTR) by targeting the liver itself. The drug doesn’t encounter the delivery problem as the drug wrapped in nanoparticles will be delivered to patients through infusion. As Alnylam Company is focused on the discovery, development and commercialization of RNA interference therapeutics for genetically defined diseases, this approval has given a huge leap for the company to go forward with the technology. The company is seen developing other RNAi-based candidate therapies that aim to treat high cholesterol, bleeding disorders, and Parkinson’s disease.

Inside the High-Stakes World of Vaccine Development

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Vaccines are widely recognized as the most effective way to fight infectious disease, a bulwark against a staggeringly diverse array of potentially pathogenic organisms looking to circumvent our defences. Unavoidable antibodies like those for flu, measles, or polio offer a suspicion that all is well and good, however it wasn’t generally in this way, and a scope of built up and rising dangers keep on presenting genuine issues. Given the physical interconnectedness of even the most remote areas with whatever is left of the world, recondite pathogens have a most optimized plan of attack to worldwide transmission more than ever.

In this context, the front lines of vaccine development research –– represent a life-and-death battle for millions of people around the world.

There are four essential players in the field of modern scale antibody innovative work: Pfizer, Merck, Sanofi, and GlaxoSmithKline (GSK). The look for an immunization starts with a solid regard for transformative history; this fight between irresistible operators and creatures has, all things considered, has been seething for a huge number of years.

By perceiving how the safe framework reacts to a specific danger, it’s conceivable to decide the idea of the coveted defensive reaction you’re planning to create. For instance, do you require antibodies to kill the rival by perceiving destructive trespassers? Or T cells that go “cytotoxic” and kill pathogens through chemical warfare? With an idea of the flavour of immune response you’re hoping to effect, the search for an antigen begins in earnest.

Antigens are sub-atomic warnings – peptides, sugars, or lipids that coat the external surface of a pathogen and raise the host cell’s alerts. There are frequently many different antigens that lead to a number of different host responses, some more effective in neutralizing and clearing the pathogen than others. It’s a torturous process to find the most useful antigen-antibody pairing (there’s a reason this takes teams of the world’s best scientists decades), but once you’ve got something promising, it’s on to the manufacturing step.

In most industries, purity is a primary goal. For vaccines, early endeavors demonstrated that evident contaminants in a cell culture – oils, salts, arbitrary natural goo – could really enhance invulnerable reaction. These supplementary molecules are called adjuvants; the search for an adjuvant that modulates the antibody-antigen response just so is another time-consuming step.

The final product is frequently a mess of atomic parts. GSK’s shingles antibody, which is as of now experiencing preliminaries, comprises of “a compound from the cell mass of Mycobacteria, another compound separated from the bark of a tree from Chile, consolidated in a liposome vesicular structure,” as indicated by Slaoui.

Despite this optimism, the hardest target is HIV. We haven’t yet uncovered where the virus lies latent, when it is absolutely stealthy. Be that as it may, figuring out how to focus on the infection in all periods of contamination and illness movement – possibly through a scope of immunization medications – isn’t inconceivable.

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CD-8 cells to fight Cancer and Chronic Infections

Immune system has several components like cytokines, lymphocytes, macrophages etc. CD-8 cell is one major component produced by the host immune system to fight pathogens like bacteria, virus etc. Upon exposure to any invading pathogen like virus, CD8 cells multiply rapidly. At the initial stage they are effector cells, acting like foot soldiers and killing the pathogen. Once the pathogen is destroyed, most of these effector cells suicide to not continue attacking the body’s own cells. Few effector CD8 cells that survive become memory cells guarding the host from the respective pathogen and enacting faster combat reaction upon exposure to same pathogen.1

CAR T cell therapy gathered lot of attention from the public for its effective use in immunotherapy against cancer and chronic infections like HIV. Application of CD-8 cells in immunotherapy has been discussed recently. The usual problem encountered is that CD8 cells get exhausted or stop functioning properly in cancer and HIV infections.  However recent research by Shomyseh Sanjabi and her team have discovered a great finding which could offer a greater option against cancer and chronic infections.

The team identified 2 molecules namely Sprouty 1 and Sprouty 2. These molecules are known to modify the survival and development of effector and memory CD8 cells respectively. Upon animal model research, the team found that in absence of these molecules in CD8 cells, the CD8 effector cells survived in larger numbers and became memory cells. The memory cells without these molecules had better protective capacity against bacterial pathogens than normal CD8 memory cells with Sprouty molecules.

In tumors, as tumor cells consume lot of glucose the effector CD8cells get killed due to glucose deprive however the CD8 cells without Sprouty 1& 2 molecules can survive and function in a tumor environment upon consuming less glucose. Also the memory CD8 cells without Sprouty molecules can tackle cancer cells and also cells activated with latent virus in viral infections. Hence the memory plays a good role in immunotherapy and the future engineering of CAR Tcells in combination with genome editing technique like CRISPR can help in future to eliminate the Sprouty molecules and employ CD8 cells against cancer and infected cells.

Are Autoimmune diseases Fatal?

If any organism produces immune response against its own healthy cells and tissue systems, then the phenomenon is known as Autoimmunity and the disease from such an aberrant immune response that results is termed an “autoimmune disease”.

Immunodeficiency and Autoimmunity:

Clinical and laboratory characteristics of autoimmunity are presented in large numbers by immunodeficiency syndromes. Autoimmunity through perpetual immune system activation may be caused to clear infections in these patients by the decreased ability of the immune system. Autoimmune thyroid disease, autoimmune thrombocytopenia and inflammatory bowel are the multiple autoimmune diseases seen from common variable immunodeficiency.

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Diagnosis:

It can be diagnosed on an accurate history and physical examination of the patient and even on certain abnormalities in routine laboratory tests. In many of the systemic disorders, specific autoantibodies can be detected from serological assays, whereas immunofluorescence of biopsy specimens is the best to diagnose localised disorders. And autoimmune diseases are diagnosed by many Autoantibodies and these levels are measured to determine the progress of the disease.

Treatments:

Autoimmune disease treatments have traditionally been anti-inflammatory, palliative, or immunosuppressive. In autoimmune diseases inflammation managing is highly critical. These are palliative treatments as the outcomes are autoaggressive responses in cases such as hormone replacement in Hashimoto’s thyroiditis or Type 1 diabetes mellitus which are non-immunological therapies. TNFα antagonists, the B cell depleting agent rituximab which are immunomodulatory therapies are shown to be useful in treating Rheumatoid arthritis which indeed these immunotherapies will be associated with increased risk to infection.

Newly Made Mitochondrial DNA Drives Inflammation

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Actuation of the inflammasome protein complex in safe cells is a key advance that triggers a natural invulnerable reaction. It rises that the blend and oxidation of mitochondrial DNA drives this enactment step. Mitochondria can control how safe cells react to contamination and tissue harm. For instance, these organelles can deliver expert or calming signals by adjusting the levels of metabolites created in the Krebs cycle or by changing the level of generation of receptive oxygen species (ROS). An ever increasing number of cases are being found of mitochondrial capacities being repurposed in startling approaches to add to incendiary signalling.

The intrinsic resistant reaction mounts a barrier when invulnerable cells perceive general signs of disease, for example, lipopolysaccharide (LPS) atoms, which are available in numerous kinds of bacterium. Nonetheless, the wrong releasing of an inborn resistant reaction can prompt immune system issue. Picking up a superior comprehension of how inborn safe reactions are managed may prompt enhancements in clinical medicines for such scatters.

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The inflammasome is a multiprotein complex that collects in invulnerable cells amid an intrinsic safe reaction. It gives cautious capacities when the inflammasome-related chemical caspase-1 separates and actuates incendiary proteins, for example, IL-1β. Inflammasomes that contain the protein NLRP3 can frame in insusceptible cells called macrophages, and the underlying strides in the get together or preparing of this sort of inflammasome are sensibly surely knew: whether LPS ties to the receptor protein TLR4 on the macrophage surface, there is an expansion in motioning by the NF-κB pathway. This causes an expansion in articulation of NLRP3 and of the antecedent type of IL-1β.3

In any case, the procedure that triggers inflammasome actuation, which happens when the compound caspase-1 is selected to the inflammasome and helps the generation of incendiary proteins, isn’t completely comprehended. It was bewildering that numerous very differing atomic signs can trigger this progression. However implies from exploratory examinations have proposed that these signals may at last act through a mitochondrial pathway related with abnormal amounts of mitochondrial ROS3,— which are required to oxidize mitochondrial DNA — and the arrival of oxidized mitochondrial DNA, which ties to the inflammasome.4

This finding of yet another fascinating link between mitochondria and inflammatory signalling in the innate immune system might reflect the organelle’s early evolutionary origins as a bacterial cell. This inherent otherness could give mitochondria a head start in being recognized as foreign by the innate immune system.

VAR2CSA DBL1-2: A Lead Antigen for Malaria Vaccine Development

12Jungle fever caused by Plasmodium constitutes a noteworthy medical issue is as yet a standout amongst the most widely recognized dangerous irresistible maladies on the planet. In 2016, the worldwide count of jungle fever achieved 216 million cases and 445,000 passing, a substantial larger part of them coming about because of Plasmodium falciparum infection.1 Individuals living in high P. falciparum transmission settings step by step obtain insusceptibility to the most serious clinical signs of the contamination. Every year, PM is in charge of 20% of stillbirths in sub-Saharan Africa, 11% of every infant passing in sub-Saharan Africa, and 10,000 maternal passing all around.

Over 50 million women are exposed to the risk of malaria during pregnancy every year. Malaria during pregnancy is a leading global cause of maternal morbidity and adverse pregnancy outcomes. Adhesion of infected erythrocytes to placental chondroitin-4-sulfate (CSA) has been linked to outcome of placental malaria. Accumulated evidence strongly supports VAR2CSA as the leading placental malaria vaccine candidate.13

Recombinant proteins encompassing the VAR2CSA high affinity CSA binding site have been generated, and their activity as immunogens that elicit functional (inhibitory) and cross-reactive antibodies against CSA-binding parasites assessed.14

The expression of His-tagged proteins was compared in four different expression systems and their capacity to bind specifically to CSA was analysed. CHO cells and E. coli Shuffle cells were the two expression systems able to express some of the recombinant proteins in reasonable amounts. Larger analytical scale production of DBL1x-2× (3D7) and DBL3x-4ε (FCR3) best expressed in CHO and E. coli Shuffle cells were performed.

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Purified proteins were administered to rats either alone or adjuvant with human approved adjuvants. Analysis of the functionality and cross-reactivity of the induced antibodies allowed us to down-select the DBL1x-2(3D7) expressed in E. coli Shuffle cells as the best antigen to be transitioned to further clinical development in order to protect future pregnant women living in malaria endemic areas against the severe clinical outcomes of placental malaria.

The Genuine behaviour of cells defamation in Alzheimer’s revealed:

Insusceptible cells usually faulted in Alzheimer’s and other neurodegenerativMicrogilae ailments are really exactness cleaning machines ensuring the focal sensory system. By valuing the part of these cells in full, researchers are better situated to grow new medications and tailor prescription to singular patients’ needs. It’s imperative to know the part and capacity of these cells, particularly going ahead for human treatment. Researchers realized that the cells assume a basic part in mental health, yet their part in adulthood was substantially murkier, with numerous researchers contending their movement was hurtful. The exploration uncovers that damage to the focal sensory system enacts the microglia, and the cells react with astounding accuracy. The analysts likewise noticed how rapidly the cells were changing and how rapidly they cleared flotsam and jetsam. Researchers taking a shot at medications for neurological ailments may need to figure that.

Another Molecular imaging strategy can screen the accomplishment of quality treatment in every aspect of the mind, conceivably enabling doctors to more successfully handle cerebrum conditions, for example, Parkinson’s infection, Alzheimer’s sickness and various sclerosis

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Therapeutic Agents:

Microglia were the disregarded cells of the mind for a considerable length of time, Researchers said that the tide is changing, and we presently acknowledge how intriguing and interesting the science of these cells is? This work demonstrates the physiological reaction of microglia after CNS [central nervous system] damage, which is altogether different from their part in neurodevelopment or in perpetual pathologies, for example, Alzheimer’s malady. Understanding microglia science in physiology and pathology will convey us closer to advancement of another age of helpful operators for neurological clutters.

Methods for Diagnosing Viral Infections

Immunoassay-based Tests:

Antibodies produced immediately after the invasion of a foreign substance can inform on primary infection, reinfection or a reactivation state. Therefore, measuring the level of immunoglobulin’s (Ig) is a widely considered approach for the diagnosis of viral infections. Immunoassays use labels conjugated to synthetic antibodies or antigens which are linked to a solid phase, and used to capture corresponding antigens or antibodies present in sera samples. These labels could be radioactive isotopes, enzymes that cause a change in colour or light-generating substances. Consequently, this principle has generated several methodologies for the testing.

Radio-immunoassay (RIA) is probably the initiating method; it uses radioisotopes to label antigen or antibody. The amount of substance to analyse is determined by the amount of the generated radioactivity. RIA is a highly sensitive method but the main drawback is the handling and disposal of hazardous radioactive substances.

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The enzymatic labelling alternative using alkaline phosphatase or horseradish peroxidase as markers is, however, the most widely used and was long considered a reference method. These enzymes induce emission of signals or change in colour respectively and allow the amount of analyst of interest to be measured. This enzyme-linked immunoassay (EIA) has numerous variants, including ELISA, and they differ in the enzyme used and the signal detection principle.

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Automated immunoassay techniques for virus detection overcome some of the limitations encountered with the conventional tests, particularly the delay to respond.

One of the limitations is that the Immunoassays are more prone to interferences than any other assay, which leads to false-positive or false-negative results.

Amplification-based Assays:

Nucleic acid amplification by polymerase chain reaction (PCR) has revolutionized the field of molecular diagnosis. The basic PCR assay relies on extraction and purification of the nucleic acid, then exponential amplification of the target sequence, using a thermostable polymerase enzyme and specific primers. The resulting amplicons are then identified using a fluorescence-based detection system, and the result is reported in international units IU/ml.

Soon after its invention, modifications in PCR were tested and patented, with the aim of improving the assay capabilities. The term nucleic acid amplification tests (NAAT) was applied to this range of new variants. NAAT is very popular in the diagnosis and management of viral infections because they allow determination of the viral load. In other terms, quantitation of the viral nucleic acid by amplifying the target sequence thousands-fold. The most widely used variants of conventional amplification are real-time PCR (quantitative PCR) and reverse transcription-PCR (RT-PCR). Both are nowadays becoming benchmarks in assessing the viral load, and while the first method quantifies DNA throughout the reactions in real time the second performs RT of the mRNA (RNA messenger) and amplifies the resulting cDNA (complementary DNA). It also quantifies RNA. The combination of both techniques increases sensitivity in detecting viruses, particularly influenza viruses. Other amplification-based tests such as nucleic acid sequence-based amplification (NASBA) and transcription-mediated amplification (TMA) are suited for detection of RNA viruses by amplification of the mRNA instead of conversion to cDNA.

The limitations of PCR are an important parameter to consider, despite the cost-effectiveness and reliability in the diagnosis of viral infections. The risk of contamination is very high while handling, especially during the sample preparation step, in addition, real-time PCR has a longer run-time (2–5 h) by comparison to other techniques. The high mutation rate of some viruses could trigger mutation within PCR primer regions of the viral genome, which would lead the virus to escape the detection by this assay.

Next Generation Sequencing:

Next-generation sequencing (NGS) is one of the greatest achievements of the modern era. Beyond genome sequencing from known organisms, it allowed discovery of novel viruses responsible for unknown human diseases, and tracking of outbreaks and pandemics such as influenza to understand their emergence and transmission profiles. Improvements and automation have dramatically increased the speed and accuracy in delivering maximum volume of data comparing to dideoxynucleotide sequencing. Technically, NGS is inclusive of three main steps: sample preparation, sequencing and data analysis. Efficient and accurate clinical diagnosis of viral infections using NGS is increasingly aiming to provide accurate longer read-length in the shortest time and at a lower cost. Bioinformatics platforms are key components of the sequencing process. They allow interpretation of the sequencing output through computational analysis and then convert it into useful information on species, genotypes and the occurrence of mutations conferring virulence or resistance to antivirals.

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NGS is undeniably a key technology in specialized clinical laboratories, but its implementation is still a challenge in many countries, where not only their resource-limited settings cannot afford a sequence analyser, sample and library preparation, but the vast majority of the population cannot afford the cost of the test.

Mass Spectrometry:

Mass spectrometry (MS) is nowadays a benchmark of laboratory qualitative and quantitative investigation, particularly in bacteriology. The principle of MS relies on converting the sample into charged particles (ions) by ionization process. These ions are separated according to their mass-to-charge ratio (m/z) and analysed by a detector. The result obtained is compared to a reference database (library), existing within the system and delivered as an interpretive spectrum.

In clinical laboratories, matrix-assisted laser desorption ionization (MALDI) and electrospray (ES) are the most used ionization methods because they allow processing of considerable amounts of analyte. The combination (RT-PCR/ESI-MS) was able to detect viral pathogens usually undetected by regular testing methods and provided rapid and detailed data (types and subtypes) within a short time. The blend of two powerful machineries (PCR-MS) can detect drug resistance to antiviral therapy as well as the presence of multiple viruses within the same sample and diagnose for co-infections.

Mass spectrometric-based methods are versatile, sensitive, rapid and cost-effective, and do not require interpretation software for data analysis. The automated machinery necessitates easy sample preparation and fewer operators. The analysis capacity can reach up to 960 specimens/day, which makes it suitable for routine diagnosis in high-volume laboratories and large-scale studies. Tests can also be performed efficiently on an archived specimen.

The main limitation of MS is the high cost, particularly in high pandemic areas, which are usually the poorest; not all laboratories can afford a mass analyser for their activities. The second major drawback is within the reference library. The identification is limited by known data from well-identified organisms only; therefore, rare mutations cannot be detected if they do not exist within the reading platform, but there is hope that MS database libraries will rapidly expand.

Boosting the Immune System to Fight Cancer

There have been cancer treatments with such a promising future as utilizing the patient’s own particular resistant framework known as Chimeric antigen receptor T-cell treatment (CAR-T) this treatment utilizes re-designed executioner T-cells to assault tumour cells, yet it likewise causes conceivably lethal symptoms.

The present CAR-T framework has three noteworthy defects: target specificity; quality of reaction; and absence of versatile capacity, which is basically the issue of backsliding. The recent system has the ability to address those three problems.

Customary CAR-T is a treatment designed for one particular patient to treat one particular sort of tumour cell. The new refined framework—called split, all-inclusive, and programmable (SUPRA) CAR-T—can be constantly adjusted to target diverse kinds of growth cells, turned on and off, and generally offers an altogether more finely tuned treatment than the present treatments.

Rather than calling them as engineering cells, it is preferable to mention them as an Antibody, which drags killer t-cells along with it. When they are bonded and activated they are able to involve more T-cells and able to make identical copies which drugs lack.

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This staggering immune reaction is additionally what causes the extreme symptoms. What’s more, there have been propelling in medicate treatment to relieve these reactions by blocking pointless segments of the insusceptible reaction while as yet enabling the CAR-T to assault the malignancy cells. What’s more, the more prominent number of disease cells implies a more grounded resistant reaction. Be that as it may, the SUPRA CAR-T framework would give specialists a chance to deactivate the whole treatment in the event that the symptoms turned out to be excessively extreme.

SUPRA CAR-T parts the T-cell from the objective detecting bit of the framework. The objective on tumour cells is called an antigen and whichever antigen is picked is searched out by an immune response on the CAR T-cells. The new framework breaks separated the T-cell from the counteracting agent and take into consideration the capacity to switch targets; similarly as unplugging a lightning link from a connector and connecting to an alternate charging link.

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Another element this split framework produces is the capacity to finely tune and enacting the T-cell reaction, which mitigates the hazardous symptoms of past CAR-T frameworks. By introducing a component that can block the binding of the T-cell and the antibody, the SUPRA CAR-T system can be deactivated. The level of deactivation can be tuned by choosing the strength to which this component binds to the antibody.

Immunosequencing Overview

Revolutionary New Breakthrough in Skincare is Light Therapy for Acne

1 Acne is a bummer. Whether you’re a teenager struggling through puberty or still fighting breakouts in your adulthood, acne can lead to feelings of insecurity and self-doubt. But don’t let blemishes keep you from facing the world with confidence. Today there are more treatment options available than ever, including light therapies that don’t require harsh topical creams that irritate or dry out your skin. Read on to learn how you can achieve great results and get the clear skin you’ve always desired.

 The skincare industry is constantly evolving. Because technology and techniques are always improving .Acne is the medical term for plugged pores of blackheads and whiteheads, pimples, and even deeper lumps of cysts or nodules that occur on the face, neck, chest, back, shoulders and even the upper arms. Pores are connected to the oil glands under your skin by a canal called a follicle. The oil from your glands carries dead skin cells to the surface of your skin and sometimes clumps together to cause a plug. The bacteria in the plug can cause swelling and forms a pimple. Though harmless and not threatening to your health, acne can cause scarring.

 For acne light therapy is an aid in preventing papules and pustules. When light is focused on skin, it penetrates into various layers and helps kill acne bacteria and reduces inflammation. Light therapy is a pain-free procedure and we can have lasting results when used on a regular basis. There are several FDA-approved products in the market to make acne treatment possible.

 The color of the visible light used in light therapy will determine how deeply it can penetrate your skin. Most dermatologist use red or blue light, or a combination of both to treat acne. Blue light is most commonly used, resulting in about a 60-70% reduction of acne. On the other hand, red light can penetrate your skin more deeply, making it able to reach sebaceous glands. When combined, red and blue light used together can maximize the benefits.