Manual Visualizing Immunity

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Visualizing the dynamics of immune surveillance in brain tumors by intravital multiphoton microscopy.

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Brain tumors BTs generally have a bad prognosis despite conventional treatment strategies. Immunotherapy is a relatively novel treatment approach that has shown benefit for durable treatment of melanoma, and is a promising candidate for different tumor types including BTs. However, little is known about the dynamics and regulation of T cell surveillance in BTs.

Resident immune cells of the myeloid lineage known as microglia are ubiquitous in the brain parenchyma while tissue-resident myeloid dendritic cells DCs known to activate T cells are relatively rare in the brain compared to DCs in other organs. Accumulating evidence indicates that myeloid cells infiltrate and create an immune suppressive microenvironment in BTs, but the identity of these myeloid cells and their role in the adaptive immune surveillance of BTs by T cells is unclear.

Based on the predominance of microglia in the brain tissue, studies focused on understanding how BT immune surveillance is regulated, have been skewed toward microglia.

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To discover the fluorogenic probe, the researchers screened a library of derivatives of a promising candidate compound for their microglia binding abilities. The best probe identified was called CDr20, which stands for "cell designation red 20", and it selectively recognized microglia in brain tissue.

Visualizing immunity in action: The extraordinary benefits of live cell imaging and analysis

The molecular base of this recognition was also studied. The fluorogenic probe contains an aromatic styryl component, which is functionalized with a hydroxy group. Without this function, the binding was not specific and no fluorescence signal was detected.

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Subsequently, the researchers looked for the exact target of the probe. Genetic studies revealed that gene expression of a protein called Ugt1a7c was linked to fluorescence signaling; thus, it was clear that enzymatic activity of this protein induced the fluorescence. The scientists found that the Ugt1a7c protein catalyzed the coupling of CDr20 with a glucuronic acid moiety, and only with this unit attached did CDr20 fluoresce.


The Ugt1a7c gene belongs to the Ugt gene family of enzymes, and the team notes that, "interestingly, in the Ugt gene family, the Ugt1a7c gene was the only member enriched in microglia. Selective microglia labeling and imaging also worked in live brains. How does it exert pressure on the genetic diversity of cancer cells?

Scientists from the Institut Pasteur and Inserm used in vivo video techniques and cell-specific staining to visualize the action of immune cells in response to the proliferation of cancer cells. The findings have been published in the journal Science Immunology on November 23, Over time, the uncontrolled proliferation of tumor cells results in the accumulation of new mutations and changes to their genome.

Intravital microscopy: visualizing immunity in context.

This gradual process creates significant genetic diversity among the cancer cells in any given patient. And although the cells in the immune system, especially T cells, are potentially able to eliminate these abnormal cells, tumor diversity can have a harmful effect, complicating the action of the immune system and rendering some therapies ineffective. Understanding this frantic race between tumor development and the immune response is key to the success of future immunotherapy techniques.